Issue 1 - Thermo Fisher

Cell Culture people you 

can contact today... 

Tina Mclure (Australia) 

Product Specialist – Cell Culture 

tina.mclure@thermofisher.com 

ph: +61 3 9757 4382 

Tina has been involved in the sales and product 

management of tissue culture products over the 

past 5 years and has just returned from visiting 

the Nunc and Nalgene manufacturing sites. Tina 

is very excited with the introduction of a number 

of new and innovative products for tissue culture 

work for 2010 and 2011. 

Peter Chisholm (Australia) 

Product Manager – Cell Culture and BioProcess 

peter.chisholm@thermofisher.com 

ph: +61 3 9757 4457 

Peter has been part of the life science industry 

for more than 30 years, and still loves what 

he is doing. Peter is always willing to share 

his industry-wide experience and expertise 

particularly in the Bioprocessing domain. 

Peter’s key responsibility is HyClone – the 

quality name in cell culture media, sera and 

bioprocessing containers. 

Jerry Wong (New Zealand) 

Product Specialist – Cell Culture 

jerry.wong@thermofisher.com 

ph: +64 9 980 6768 

Jerry completed his Bachelor of Technology 

honours degree (Biomedical Science) at the 

University of Auckland, and worked at Vialactia 

Biosciences Ltd before embarking on completing 

his PhD at the University of Auckland. Jerry’s 

research was in molecular neuroscience to 

investigate novel therapeutic strategies for 

Parkinson’s disease. Jerry joined our team early 

this year and will bring his extensive knowledge 

to our cell culture team. 

The Thermo Fisher Scientific Story 

Australia & New Zealand 

Thermo Fisher Scientific is the world leader in serving 

science, enabling our customers to make the world 

healthier, cleaner and safer. In April 2009 Thermo Fisher 

Scientific made a major investment in the Australasian 

region acquiring the distribution company Biolab which 

in itself has a 150 year history. 

By doing so, Thermo Fisher has taken the first steps 

to bringing together worldwide leading products and 

brands into the one company. With our continued 

commitment and investment into R&D, analysis, 

discovery and diagnostics, Thermo Fisher Scientific 

today is leading the way with an unparalleled product 

portfolio and service offering, combined with a 

customer-focused technical sales capability. 

Globally, Thermo Fisher Scientific has 33,000 

employees from 38 countries all sharing in the same 

mission and core values expressed as the four I’s: 

Integrity, Intensity, Innovation, and Involvement. So it 

is a great pleasure to have the opportunity to relaunch 

the previously known publication Helix, with the aim 

of sharing one of those key values by renaming the 

publication Bio-Innovation. 

Bio-Innovation will be published quarterly and will 

provide researchers with the latest updates on new 

applications and methodology (including new products 

and technologies) with the first issue focusing on 

Cell Culture. This issue will also be available at the 

forthcoming Australian Society of Immunology 

meeting, Perth, 5-9 December 2010. Our next issue 

will be focussed on molecular biology. 

Christopher Hum 

Business Development Manager 

Biosciences 

2

Application Note 

Contents 

Articles 

Outstanding Observation: A Faster Way To Publish Your Groundbreaking Findings 

Stem Cell Promise–Research Brings Autograft Revolution Closer 

Choosing the Right Centrifuge for Your Application 

4 

20 

22 

37°C 

Applications 

Technical Bulletin: Cell Adhesion & Growth 

5 

20°C 

UpCell Surface versus Trypsinisation and Scraping in Cell Detachment 

Take Your Cell-Based Assays To The Edge / Vita Means Life 

A Novel Feeder-Free Embryonic Stem Cell Culture System 

Endothelial Progenitors Encapsulated In Bioartificial Niches Are 

Insulated From Systemic Cytotoxicity & Are Angiogenesis Competent 

Real World Advantages In Breast Cancer Analysis 

Water Quality Standards For Research And Analysis Applications 

Testing The Efficacy Of The Antimicrobial Treatment – A Study 

Preventing Cell Culture Contamination with Copper CO 2 

Incubators 

6 

7 

8 

9 

10 

11 

18 

19 

Advancing Cell Culture 12-17 

News, Events & Exhibitions 

The Next Generation In Automated Liquid Handling 

When is a μL not a μL? 

Chart MVE for Ultra Cold Storage 

Culture Serum Q & A 

24 

25 

26 

27 

3

article 

Outstanding Observation: A faster way to publish 

your groundbreaking findings 

It is often difficult to publish novel findings if they do 

not also include a detailed description of the molecular 

mechanisms involved. Researchers may find that the 

dissemination of their groundbreaking discoveries is 

delayed by months or even years as they try to provide 

the in-depth mechanistic data required to satisfy the 

submission criteria for their desired journal. 

Professor Chris R Parish, Editor-in- 

Chief, Immunology and Cell Biology 

In an effort to break through this 

barrier and to allow immunologists 

to publish their paradigm-shifting 

results without getting held up 

with the mechanistic basis of the 

observations, Immunology and Cell 

Biology launched the Outstanding 

Observation series in 2008. 

Owing to the cutting-edge nature 

of this type of article, papers are 

expedited through the refereeing 

process and are therefore published 

more rapidly than standard articles. 

I would like to invite you to sample 2 

recent examples of this article type: 

Polyclonal Treg cells enhance the 

activity of a mucosal adjuvant 

Silvia Vendetti, Todd S Davidson, 

Filippo Veglia, et al.Immunology and 

Cell Biology (2010) 88, 698-706 

CD69 limits early inflammatory 

diseases associated with 

immune response to Listeria 

monocytogenes infection Javier 

Vega-Ramos, Elisenda Alari-Pahissa, 

Juana del Valle, et al. Immunology 

and Cell Biology (2010) 88, 707-715 

Access these articles free of charge via the 

Scan these tags with 

your mobile phone for 

instant access to 

these articles. 

Download the free app 

for your phone at 

http://gettag.mobi 

Immunology & Cell Biology website. www.nature.com/icb 

Congratulations to Dr Cindy Ma, Immunology and Inflammation, Garvan Institute of Medical Research, 

NSW, Australia, winner of the 2009 Outstanding Observation ICB Publication of the Year award. 

The award is a AU$1000 scholarship provided by the Nature Publishing Group. 

Also Congratulations to Dr Mark Dowling, Immunology, Walter and Eliza Hall Institute, VIC, Australia 

as runner-up and will receive AU$500 travel scholarship provided by Thermo Fisher Scientific. 

4

Technical Bulletin: Cell Adhesion & Growth. A comparison 

of Coated, Modified Glass and Plastic Surfaces 

Wendy K. Scholz, Thermo Fisher Scientific 


Introduction 

Historically, glass has been used as the growth 

surface for cells since it has superior optical qualities 

and is naturally charged. Disposable plastic, especially 

polystyrene is now commonly used for cell culture growth. 

Plastic culture vessels are of good optical quality, and the 

growth surface is flat. However, since most plastics are 

hydrophobic and unsuitable for cell growth, they are often 

treated with radiation, chemicals or electric ion discharge to 

generate a charged, hydrophilic surface. Such treatment of 

plastic generates a surface preferred 

over glass by many cell types. 

The growth of various cell types on plastic, soda lime glass 

and borosilicate coverglass was examined. The surfaces 

were either unmodified, coated with polylysine, or stably 

surface modified with non-biological reagents or electrical 

discharge. Glass surfaces were chemically modified in 

two ways: using a proprietary procedure and reagents 

or as described by Kleinfeld, et al. (1988). All substrates 

were assembled into Thermo Scientific Nunc Lab-Tek and 

Lab-Tek II Chamber Slide products. 

General 

Growth substrates may affect the morphology, 

differentiation and behavior of various cell types. 

A cell’s repertoire on various surfaces is cell type specific. 

Epithelial and fibroblast cell lines remain proliferative on cell 

culture treated plastic. Biological coatings and chemically 

modified surfaces may reduce the proliferation rate by 

inducing differentiation into a more mature state. Cells in 

a more differentiated state usually function and express 

proteins characteristic of the tissue of origin. 

Culturing neurons is a particular challenge since they do not 

continue proliferating after dissociation. Neuron survival in 

culture is dependent on cell adhesion and differentiation, 

which can be facilitated by modifying the surface with a 

biological coating or chemical modification.This bulletin 

examines the correlation between primary neuron adhesion 

and differentiation, and surface properties such as surface 

energy and available specific substrate molecules. 

Selection of a growth surface for cell culture 

should also be based on application. While glass and 

plastic growth surfaces are flat and optically clear they 

differ in many ways that may affect performance in various 

applications. Glass surfaces offer optimal optical clarity 

with a minimal autofluorescence and are preferred for 

most fluorescent applications. Polystyrene can be used for 

fluorescein if the proper blocking filters are used in the UV/ 

blue range and perform well at longer wavelengths. Glass 

surfaces are more resistant than plastics to solvents, acids, 

bases and heat. Fixative compatibility studies have been 

performed on glass and plastic. 

Results: 

Cells of the fibroblast-like cell line BHK-21 (baby hamster 

kidney), which are very adherent, did not distinguish 

between these growth surfaces. The less adherent 

fibroblast-like cell line L929 (mouse lung) and two epitheliallike 

cell lines, HEp-2 (human epidermal) and WISH (human 

amnion), grew to slightly higher densities and produced 

more uniform monolayers when grown on electrically 

modified plastic compared to unmodified plastic or glass. 

These cell types also grew very well on both types of 

chemically modified glass surfaces.Primary brain neurons 

did not adhere to unmodified plastic or glass surfaces. 

However, neurons adhered and differentiated on polylysine 

coated glass or plastic surfaces and, albeit differently, on 

both of the chemically modified glass surfaces. Electrical 

modification of the plastic or glass, which significantly 

increased the surface energy or wettability, did not produce 

a surface suitable for these neurons. Adhesion, growth and 

differentiation of cells on a surface is cell type specific and 

involves more than one mechanism. Many cell lines prefer 

surfaces with a high surface energy such as produced by 

electrical modification. Other cell types, such as primary 

neurons, require a specific interaction with functional 

groups provided by polylysine coating or chemical 

modification of the growth surface. 

Conclusions 

The effect of growth surfaces on adhesion, growth and 

differentiation of cells is cell-type specific and involves more 

than one mechanism. 

• Many cells prefer surfaces with high surface 

energies (i.e. hydrophilic surfaces). 

• Growth surfaces such as CC2 may improve cell 

adhesion and induce cellular differentiation 

of fibroblastic-like cells. 

• High surface energy is not sufficient for 

primary neuron growth. 

• Biological coatings or chemical modifications 

that place amines on the growth surface facilitate 

primary neuron adhesion and differentiation. 

• Other variables, such as those that differentiate 

European and American borosilicate coverglass, 

may influence neuron survival on glass surfaces. 

For more information or to obtain a full copy of 

this technical bulletin please contact: 

Tina McLure(AU) 

tina.mclure@thermofisher.com 

Ph: +61 3 9757 4382 

Dr Jerry Wong (NZ) 

jerry.wong@thermofisher.com 

Ph: +64 9 980 6768 

5


UpCell Surface versus Trypsinisation 

and Scraping in Cell Detachment 

This compares the recovery of mouse peritoneal 

macrophages harvested from the UpCell Surface 

using temperature reduction with those harvested 

from traditional cultureware (tissue culture-treated 

polystyrene) using trypsinisation or scraping. 

37°C 

20°C 

Methods 

Mouse peritoneal macrophages in RPMI 1640 medium 

supplemented with 10% fetal bovine serum (FBS) were 

seeded in one dish with UpCell Surface and two traditional 

cultureware dishes at 2.4 x 10 5 cells/cm 2 . The cells were 

incubated at 37°C in a humidified atmosphere of 5% CO 2 

in air. After 2 hours of incubation, non-adherent cells were 

removed by washing with phosphate-buffered saline (PBS). 

Cells were then cultured for 2 days in RPMI 1640 medium 

supplemented with 10% FBS and harvested using one of the 

following procedures: 

Harvest of cells from the UpCell Surface using 

temperature reduction 

• Non-adherent cells were removed by washing with 

Ca 2+ – and Mg 2+ –free PBS 

• 4.0 mL RPMI 1640 medium supplemented with 

10% FBS was added and the dish was incubated 

at 20°C for 30 minutes 

• Detached cells were harvested 

Harvest of cells from traditional cultureware using 

trypsinisation 



• 1.0 mL of 0.25% trypsin/EDTA was added, and the 

dish was incubated at 37°C for 5 minutes 

• 3.0 mL RPMI 1640 medium supplemented with 

10% FBS was added 

• Detached cells were harvested 

Harvest of cells from traditional cultureware using EDTA 

and scraping 



• 4.0 mL of 2.5 mM EDTA/PBS was added and the dish 

was incubated on ice for 20 minutes 

• The cells were detached by scraping and harvested 

Results 

The harvested cells were counted and the recovery 

ratio was calculated. 

Figure 1. Photomicrographs of mouse peritoneal macrophages 

on the UpCell Surface before (a) and after (b) temperature 

reduction. After temperature reduction, the cells detached from 

the surface and became spherical. After harvesting of the cells 

by pipetting, only a few cells remained on the UpCell Surface (c). 

Recovery ratio (%) 

a B C 

100 

80 

60 

40 

20 

0 

temperature Temperature reductionS Reduction Scraping Trypsinisation 

trypsinization 

Upcell Surface 

traditional cultureware 

Figure 2. Recovery ratio of mouse peritoneal macrophages 

harvested from the UpCell Surface was compared with 

recovery ratios of these cells harvested by either enzymatic 

(trypsinization) or mechanical (scraping) methods. The recovery 

of cells from the UpCell Surface was significantly higher than 

the recovery of cells harvested from traditional cultureware by 

trypsinisation or scraping. Mean and SD is shown. 

For more information please contact: Tina McLure (AU) Email: tina.mclure@thermofisher.com or Ph: +61 3 9757 4382 

Dr Jerry Wong (NZ) jerry.wong@thermofisher.com or Ph: +64 9 980 6768 

6

Take Your Cell-based Assays to the Edge 


In cell culture, a volume loss as 

small as 10% concentrates media 

components and metabolites 

enough to alter cell physiology, in 

some cases, severely. With this in 

mind, Thermo Scientific recently 

unveiled its latest advancement in 

plates for cell culture. 

The Nunc Edge 96-well plates for 

cell-based assays incorporate large 

perimeter evaporative buffer zones 

that eliminate well-to-well variability, 

while dramatically reducing the 

overall plate evaporation rate to 

lower that 2% after seven days 

of incubation. 

And what’s the result? Tests have 

shown more viable and healthy 

cell yields are obtained. 

The unique buffer zones enable 

the use of all 96 wells on the plate 

and greatly reduce the edge effect 

commonly experienced in cell culture 

and maintaining data consistency 

throughout the plate. 

The implications of volume loss can 

concentrate media components 

and metabolites enough to alter cell 

physiology, a phenomena which is 

more pronounced in the outer and 

corner wells. The Nunc Edge plates 

significantly reduce this occurrence, 

effectively maintaining sample 

concentrations over long periods of 

incubation. The prevention of cell 

death and toxicity in the plates’ outer 

wells allows results to remain more 

true to the population phenotype 

for more efficient, high throughput 

analysis. 

In cell-based assays where multiple 

targets are simultaneously imaged, 

all the fluorescence from a single 

location needs to focus at the same 

point within the imaging system. The 

flatness of the plates eliminates the 

occurrence of chromatic aberrations, 

enabling efficient automated 

imaging. Furthermore, varying 

reagent concentrations from assay 

washing/aspiration steps are also 

dramatically reduced. 

Advantages of Thermo Scientific 

Nunc Edge Plate 

• Low-evaporation “moat” 

• 96 well format - follows ANSI standard 

footprint and is amenable to automation 

• Simply fill moat with sterile H 2 

O and 

eliminate edge effects 

• Cell culture treated or non-treated 

• Low fluorescence 

• Custom barcoding available – the safest 

way to track your samples 

• Designed for both automated 

or manual use 

• Superior imaging 

For more information please contact: 

Tina McLure (AU) tina.mclure@thermofisher.com 

or Ph: +61 3 9757 4382 

Dr Jerry Wong (NZ) jerry.wong@thermofisher.com 

or Ph: +64 9 980 6768 

Vita means Life : NEW Thermo Scientific 

Nunc Nunclon Vita Surface 

Animal component-free surface for growth of stem cells & other fastidious cells. 

Nunclon Vita Surface supports 

attachment, colony formation and 

growth of human ESC, human IPS 

cells and other fastidious cell lines in 

the absence of feeder cells and matrix 

coatings. In media supplemented with 

ROCK inhibitor, human ESC can be 

cultured on the Nunclon Vita Surface 

for at least 10 passages without loss 

of pluripotency. Human ESC has 

successfully been expanded for more 

than 10 passages without changes to 

the karyotype. 

The unique Nunclon 

Vita surface helps 

remove variability, requires 

less work, and allows for the 

treatment surface to be adaptable 

to scalable cell expansion. 

Human Embryonic Stem Cells 

(human ESC) 

• No matrix coating or feeder cells necessary 

• Expand for more than 10 passages in conditioned 

media with a ROCK inhibitor while maintaining pluripotency 

• Passage with enzymes or by mechanical selection 

• Alternatively, passage by withdrawing ROCK inhibitor followed by 

a short incubation and gentle pipetting 

7


A Novel Feeder-Free Embryonic Stem Cell Culture System that 

Supports Mouse Embryonic Stem Cell Growth and Proliferation 

Kalle Johnson 1 , Robin Wesselschmidt 2 , Mark Wight1, Thomas I. Zarembinski 3 

Figure 1: Phase contrast 

photographs of mESC 

colonies over the course 

of the study. Photos 

of colonies grown on 

HyStem-C (top Row) 

and photos of colonies 

co-cultured with MEF 

feeders (bottom row). 

Morphologies bear 

slight differences 

between HyStem-C 

and MEF co-cultures. 

However, cultures grown 

on both substrates were 

able to maintain colonies 

that generally exhibited 

relatively regular, phasebright, 

well defined 

borders, characteristic of 

undifferentiated mESC 

colonies. 

HyStem 

iMEF 

Passage 1 

Passage 2 Passage 3 Passage 4 Passage 5 

1 

Thermo Fisher Scientific 

Inc., 925 West 1800 

South, Logan, Utah 

84321 

2 

Primogenix Inc., 165 

Missouri Blvd, Laurie, 

MO 65038 

3 

Glycosan BioSystems, 

Inc., 675 Arapeen Drive, 

Suite 302, Salt Lake City, 

Utah 84108 

References: 

1) Gerecht S et al, 

Hyaluronic acid hydrogel 

for controlled 

self-renewal 

and differentiation of 

human embryonic stem 

cells PNAS 2007 vol 104: 

11298–11303. 

2) Engler et al, Matrix 

Elasticity Directs 

Stem Cell Lineage 

Specification Cell 2006 

vol 126: 677-689. 

For more information 

please contact: 

Peter Chisholm (AU) 

peter.chisholm@ 

thermofisher.com 

Ph: +61 3 9757 4457 

Jerry Wong (NZ) 

jerry.wong@ 


Ph: +64 9 980 6768 

Standard embryonic stem cell culture systems require 

co-culture with mitotically inactive mouse embryonic 

fibroblast feeder cells (iMEFs) to maintain their 

undifferentiated proliferative state. 

While iMEFs provide a suitable attachment surface 

and crucial soluble factors promoting embryonic stem 

cell (ESC) growth and proliferation, iMEFs are timeconsuming 

to prepare. More importantly, iMEFs vary 

from lot-to-lot and contaminate ESCs with carry over 

iMEFs from previous culture passages. 

These latter shortcomings confound basic research 

attempting to dissect the culture components or 

underlying gene and protein expression patterns 

important for ESC proliferation and differentiation. 

Development of a feeder-free system for embryonic 

stem cell culture using a synthetic matrix would provide a 

ready-to-use substrate which is consistent from lot to lot 

without iMEF carry over. 

iMEFs make abundant amounts of hyaluronic acid (HA) 

which is important for not only embryogenesis but 

also for human embryonic stem cell culture (1) . Using 

mouse embryonic stem cells (mESCs) as a model 

system, we reasoned that the use of a commercially 

available HA-rich matrix would provide a suitable starting 

point for preparing a novel feeder-free substrate for 

undifferentiated growth. 

Here we report the use of a crosslinkable HA-based 

substrate (HyStem-CTM) for feeder-free propagation 

of mESCs in the presence of FBS. mESCs plated on 

HyStem-C maintain excellent morphology and offer 

comparable plating efficiency to those grown on iMEFs. 

Data from FACS analysis as well as immunocytochemistry 

to confirm the presence of key recognised pluripotency 

markers will be presented. (Figure 1) 

This novel feeder-free cell culture system has potential 

uses in proteomic analysis since no carryover iMEF 

proteins will be present in embryonic stem cell extracts. 

In addition, statistics from high content analysis and 

high-throughput screening efforts employing mouse 

embryonic stem cells will be improved due to increased 

consistency during the screening campaign. 

Finally, the possibility exists to alter the stiffness or 

composition of the matrix in a manner that may enhance 

efforts to drive the pluripotent cells down desired 

lineages (2) . 

Three types available 

• HyStem Hydrogel - Chemically modified 

hyaluronan crosslinked with PEFDA 

• HyStem-C – HyStem with chemically 

modified gelatin added 

• HyStem-HP – HyStem-C with 

chemically modified heparin 

Hydrogels can be easily customised 

• by adding ECM proteins 

• by varying the Hydrogel compliance to match the 

stiffness of the native tissues 

• easy control over the amount and type of ECM protein 

incorporated 

• no autofluorescence 

• ideal for non-adherent cells – easy to mobilise 

• for adherent cells, simply add choice/selective cell 

attachment peptide 

• allows encapsulation and implantation of cells 

8

Endothelial progenitors encapsulated in bioartificial niches are 

insulated from systemic cytotoxicity & are angiogenesis competent 

B. B. Ratliff, 1 T. Ghaly,1* P. Brudnicki, 1 * K. Yasuda, 1 M. Rajdev, 1 M. Bank,1 J. Mares, 1 A. K. Hatzopoulos, 2 and M. S. Goligorsky 1 

Special Interest 

Endothelial progenitors encapsulated in bioartificial 

niches are insulated from systemic cytotoxicity and 

are angiogenesis competent. First published April 21 st , 

2010—intrinsic stem cells (SC) participate in tissue 

remodeling and regeneration in various diseases and 

following toxic insults. Failure of tissue regeneration is in 

part attributed to lack of SC protection from toxic stress of 

noxious stimuli, thus prompting intense research efforts 

to develop strategies for SC protection and functional 

preservation for in vivo delivery. One strategy is creation 

of artificial SC niches in an attempt to mimic the 

requirements of endogenous SC niches by generating 

scaffolds with properties of extracellular matrix. Here, 

we investigated the use of hyaluronic acid (HA) hydrogels 

as an artificial SC niche and examined regenerative 

capabilities of encapsulated embryonic endothelial 

progenitor cells (eEPC) in three different in vivo models. 

Hydrogel encapsulated eEPC demonstrated improved 

resistance to toxic insult (adriamycin) in vitro, thus 

prompting in vivo studies. Implantation of HA hydrogels 

containing eEPC to mice with adriamycin nephropathy 

or renal ischemia resulted in eEPC mobilization to 

injured kidneys (and to a lesser extent to the spleen) 

and improvement of renal function, which was equal or 

superior to adoptively transferred EPC by intravenous 

infusion. In mice with hindlimb ischemia, EPC 

encapsulated in HA hydrogels dramatically accelerated 

the recovery of collateral circulation with the efficacy 

superior to intravenous infusion of EPC. In conclusion, HA 

hydrogels protect eEPC against adriamycin cytotoxicity 

and implantation of eEPC encapsulated in HA hydrogels 

supports renal regeneration in ischemic and cytotoxic 

(adriamycin) nephropathy and neovascularization of 

ischemic hindlimb, thus establishing their functional 

competence and superior capabilities to deliver stem 

cells stored in and released from this bioartificial niche. 

INTRINSIC ADULT STEM CELLS participate in tissue 

remodeling and regeneration in various chronic diseases 

and following noxious cardio- and/or nephrotoxic insults 

(30). Failure of tissue regeneration is in part attributed 

to the fact that stem cells are not entirely protected 

from cytotoxic or genotoxic stress of noxious stimuli. 

One of the striking examples of stem cell vulnerability is 

represented by the toxicity profile of a commonly used 

chemotherapeutic agent doxorubicin (adriamycin). This 

agent leads to severe cardiomyopathy and nephropathy 

due to induction of oxidative stress, and stem cells 

are equally affected. Similar vulnerability of stem cells 

has been reported in other chronic diseases, such as 

atherosclerosis, essential hypertension, preeclampsia, 

hyperglycemia, smoking, and type I and type II diabetes, 

to name a few. Inexorable progression of these diseases 

may be in part explained by the reduced ability of 

stem cells to participate in tissue regeneration. The 

efficacy of stem cell transplantation in some disease 

states is the ultimate proof of the concept that stem 

cell incompetence developing in the course of chronic 

diseases and after exposure to noxious stimuli results in 

failed regeneration and progressive tissue degeneration. 

Intense research efforts have been mounted to develop 

strategies for stem cell protection and functional 

preservation. One direction is based on creation of 

artificial stem cell niches. The concept of stem cell 

niche is 30 years old yet adult stem cell niches remain 

elusive. The best-studied niches are represented by 

those harboring germline stem cells, bone marrow 

hematopoietic stem cells, and stem cells of intestinal 

crypts, to name a few. Consensus has been reached 

that the stem cell niche provides an umbrella 

microenvironment supporting cell attachment and 

quiescence by sheltering stem cells from proliferation 

and differentiation signals, enhances cell survival, 

regulates stem cell division and renewal, and coordinates 

the population of resident stem cells to meet the actual 

requirements of an organ. Creation of artificial stem cell 

niches represents an attempt to mimic some of these 

requirements by providing cells with a low-oxygen 

environment within avascular scaffolds, but ensuring 

their ability to preserve the phenotype, quiescence, 

recruitability, and protection from the noxious stimuli. 

Endothelial progenitor cells (EPC) have been shown to 

participate in regenerative processes. Transplantation of 

EPC augments neovascularization of ischemic/infarcted 

myocardium, ischemic limbs, or brain. EPC may play a 

critical role in the maintenance of integrity of vascular 

endothelium and in its repair after injury or inflammation. 

EPC are subjected to various stressors, similar to all 

somatic cells that could impair their competence. 

Hyperglycemia has been reported to reduce survival and 

impair function of circulating EPC. There is emerging 

evidence that senescence may serve as an important 

mechanism mediating EPC dysfunction. Decreased 

numbers and increased proportion of senescent EPC 

have been reported in patients with preeclampsia or 

hypertension. Angiotensin II can induce EPC senescence 

through the induction of oxidative stress and influence 

telomerase activity. Oxidized low-density lipoprotein 

induces EPC senescence and dysfunction. In addition, 

EPC dysfunction has been documented in type I and 

II diabetes, coronary artery disease, atherosclerosis, 

vasculitis with kidney involvement, and end-stage renal 

disease. Therefore, the goals of studies presented herein 

were to expand the knowledge on EPC behavior in 

hyaluronic acid (HA) hydrogels, examine the resistance of 

thus stored stem cells to noxious effects of adriamycin, 

the efficacy of EPC mobilization, and the competence of 

encapsulated endothelial progenitors to contribute to the 

postischemic organ repair and neovascularization. 

1 

Departments of 

Medicine, Physiology, 

and Pharmacology, New 

York Medical College, 

Valhalla, New York; and 

2 

Department of Medicine, 

Division of Cardiovascular 

Medicine, and 

Department of Cell and 

Developmental Biology, 

Vanderbilt University, 

Nashville, Tennessee 

For the full article 

please contact: 

Peter Chisholm (AU), 

peter.chisholm@ 


Ph: +61 3 9757 4457 

Jerry Wong (NZ) 

jerry.wong@ 


Ph: +64 9 980 6768 

9

Latest news 

Real world advantages in breast cancer analysis 

Allen M. Gown, M.D. Medical Director and Chief PathologistPhenoPath Laboratories-Seattle, WA 

Figure 1 (top right) : SP1 

(Thermo Scientific rabbit 

monoclonal antibodies) 

Figure 2: (below right) SP3 

(Thermo Scientific rabbit 

monoclonal antibodies) 

References: 

1. 

Huang Z., at el Appl 

Immunohistochem Mol 

Morphol. 2005; 13: 91-95. 

2. 

Gown AM., at el. J Clin 

Oncol. 2006; 24: 5626-7. 

3. 

Gown AM., at el. Mod 

Pathol. 2008. 

Two Thermo Scientific rabbit monoclonal 

antibodies are poised to have a significant 

impact on the immunohistochemical analysis 

of prognostic and predictive breast cancer 

markers: Thermo Scientific SP1, a rabbit 

monoclonal antibody directed against the 

estrogen receptor alpha molecule, and target 

of tamoxifen; and Thermo Scientific SP3, a 

rabbit monoclonal antibody directed against 

the HER2 transmembrane receptor, the target 

of trastuzumab (Herceptin). 

SP1 has been demonstrated to have an eight fold higher 

affinity for the estrogen receptor compared with the 

1D5 mouse monoclonal antibody that has been widely 

used in immunohistochemical analyses of breast 

cancer. 1 This higher affinity translates into a more robust 

immunohistochemical reagent, as was demonstrated 

in the paper published by Cheang et al, 2 describing a 

collaborative study performed by the British Columbia 

Cancer Agency and PhenoPath Laboratories. 

In this tissue microarray-based study of 4,150 patients 

in which determination of ER status with SP1 was 

compared with 1D5, with a median follow-up, of 12.4 

years, SP1 was found in multivariate analyses to be a 

better independent prognostic factor than 1D5. 

Furthermore, determination of ER status using the 

SP1 antibody was more precise compared with the 

1D5 antibody. The cohort, corresponding to 8% of the 

patients, who were SP1+ and 1D5-, i.e., who would have 

been classified as negative based on 1D5, were found 

to have a good outcome indicative of ER positive breast 

cancer. SP1-determined ER status also correlated better 

with ligand binding ER assay results. The study concluded 

that SP1 may represent an improved standard for ER 

assessment by immunohistochemistry in breast cancer. 

More recent studies performed at PhenoPath 

Laboratories and presented this past spring at the USCAP 

meeting in Denver 3 document the potential advantages 

of SP3 as an immunohistochemical reagent in the 

assessment of HER2 status. In a series of 421 breast 

cancers analyzed for HER2 by immunohistochemistry, 

comparing the SP3 rabbit monoclonal antibody with a 

rabbit polyclonal antibody (Dako A0485), SP3 was found 

to be a more robust reagent, producing more consistent 

run-to-run immunostaining with fewer run failures. 

The study also showed that while both antibodies 

produced results that were greater than 95% concordant 

with those of FISH, the SP3 antibody was more 

“efficient” in yielding fewer 2+ cases. SP1 and SP3 will 

undoubtedly be the subject of future studies, but the data 

to date suggest that both could well become the new 

gold standard for immunohistochemical analysis of breast 

cancer markers. 

For further information in Australia & 

New Zealand please contact: 

Julie Bloem, Business Development Manager 

Ph: +61 418 385 101 or julie.bloem@thermofisher.com 

Heidi Farrow, Product Specialist 

Ph: +61 407 844 114 or heidi.farrow@thermofisher.com 

10

Cell Culture 

Testing the efficacy of the antimicrobial 

treatment – a study 

ELGA LabWater–Water quality standards 

for Research and analysis applications 

Advancing 

LCP Research Group, Thermo Fisher Scientific, Vantaa, Finland 

Type 1+ – Goes beyond the purity requirements 

of Type 1 water’ 

Microbes, such as bacteria, fungi and algae, are found 

everywhere around us, and they are also present in the 

human skin. Normally they are not harmful, but in some 

cases they may cause deterioration of the material 

they grow on or cause cross contamination. Even 

when strict cleanliness is observed, microbes from 

the hands may contaminate any surface. Antimicrobial 

treatment protects from microbial growth and adds 

additional protection against cross- contamination. 

i.e. bacteria from a pipette that could contaminate the 

sample. 

How does it work? 

The active ingredient of the antimicrobial material 

tested is silver in the form of silver ions. In a humid 

environment the ions are slowly released from the 

inorganic matrix via an ion-exchange mechanism. 

The release is slow, but fast enough to maintain an 

effective concentration on the surface of the material. 

Silver ions are taken up by microbial cells and interrupt 

critical functions, such as DNA replication, resulting 

in the death of the microbes. The antimicrobial effect 

of the material used is longterm and silver inhibits the 

growth of a broad spectrum of microorganisms. 

Testing the efficacy of the antimicrobial treatment 

The antimicrobial effect of the material was evaluated 

according to ASTM standard E21 80. The standard 

describes a test method to evaluate (quantitatively) the 

antimicrobial effectiveness of agents incorporated or 

bound into or onto mainly flat hydrophobic or polymeric 

surfaces. The test organisms used were Escherichia 

coli, Staphylococcus aureus, Candida albicans and 

conidiospores of Aspergillus niger. 

Contamination of the antimicrobial polymer pieces 

(from a Thermo Scientific Finnpipette F1 - The handle 

and the dispensing button are made of an antimicrobial 

polymer) was carried out by pipetting 0.2 ml of the 

cell or conidiospore suspension on test pieces that 

were stored in a horizontal position throughout 

the experiments. After complete drying of the 

suspensions, the amount of colony forming units (cfu, 

a measure for viable cells) was determined after 4 

hours and after 24 hours. 

After 4 hours, a reduction of cfu was seen for all four 

test organisms. After 24 hours, the reduction was 

improved for each microorganism, except in those 

cases where 100% reduction was already achieved at 

the 4 hour mark, see Figure 1. 

These results show that the antimicrobial material 

results in a significant reduction of microorganisms, 

demonstrating the efficacy of the antimicrobial 

polymer. 

Please note: The antimicrobial treatment does not 

remove dirt and does not protect users or others 

against bacteria, viruses or other disease organisms. 

Figure 1. Reduction of model microorganisms on an 

antimicrobial polymer. The colony forming units were 

determined at 4 and 24 hours after inoculation 

Scientists perform a vast range of 

applications in many different kinds 

of laboratories. Therefore, different 

grades of water must be purified 

and utilised to match the required 

procedures or appliances. Water is 

one of the major components in many 

applications, but the significance of 

its purity is often not recognised. 

In this section we highlight some common applications 

and provide guidance on the water quality required. 

We also provide some guidance on what purification 

technologies you should be looking for in your water 

system. There are many water quality standards 

published throughout the world, however only a few 

are relevant to specific research applications. This has 

resulted in the majority of water purification companies, 

including ELGA, adopting broad generic classifications 

defined by measurable physical and chemical limits. 

Throughout this application note we will refer to the 

“Types” of water referred to in this chart (see right). 

Type I – Often referred to as ultra pure, this grade is 

required for some of the most water-critical 

applications such as HPLC (High Performance Liquid 

Chromatography) mobile phase preparation, as well 

as, blanks and sample dilution for other key analytical 

techniques; such as GC (Gas Chromatography), 

AAS (Atomic Absorption Spectrophotometry) 

and ICP-MS (Inductively Coupled Plasma Mass 

Spectrometry). Type I is also required for molecular 

biology applications as well as mammalian cell 

culture and IVF (In vitro Fertilisation). 

Type II – Is the grade for general laboratory 

applications. This may include media preparation, 

pH solutions and buffers and for certain clinical 

analysers. It is also common for Type II systems 

to be used as a feed to a Type I system*. 

Type II+ – Is the grade for general laboratory 

applications requiring higher inorganic purity. 

Type III – Is the grade recommended for 

non-critical work which may include glassware 

rinsing, water baths, autoclave and disinfector 

feed as well as environmental chambers and 

plant growth rooms. These systems can also be 

used to feed Type I systems* 

Resitivity TOC(PPB) Bacteria Endotoxins 

(MΩ-cm) 

(EU/ml) 

Type I+ 18.2

Growth and Passage 

Culture and Experimentation 

Characterisation and Analysis 

Reagent and Sample Storage 

Biological Safety Cabinets 

and Clean Benches 

Class II, Type A2 biological 

safety cabinets that provide 

best-in-class safety, 

ergonomics and energy 

efficiency for today’s most 

demanding laboratory 

applications. Unique 

technologies ensure safe 

air filtration, avoiding 

sample contamination or 

environmental hazards. 

Cell culture systems 

Our Nunc OptiCell 

cell culture system 

simplifies cell growth, 

monitoring and transport, 

while providing optimal 

conditions. 

C02 incubators 

Carbon dioxide incubators 

providing in-vitro 

environments with proven 

contamination control 

solutions. Assure that 

valuable cultures are 

secured, protected and 

thriving, delivering optimal 

growth and security. 

Filtration Products 

Filtration products 

including bottle-top 

filters, cut-disk filters, 

filter units and syringe 

filters. Rapid sterile 

filtration of even the most 

difficult to filter fluids. 

Centrifuges 

From benchtops and 

microcentrifuges to 

ultracentrifuges and 

superspeeds to carbon 

fibre rotors. Achieve 

unmatched productivity, 

versatility, performance 

and ease-of-use with our 

innovative centrifuges, 

rotors and accessories. 

High Content Screening & Analysis 

Automated imaging instrumentation, 

image analysis software, High 

Content Informatics, reagent kits, 

and screening assays and cell lines. 

Fully integrated solutions to improve 

the quality and productivity of cellbased 

assays for researchers in drug 

discovery and systems biology. 

Chart MVE 

For all your long term cryo 

storage (vapour and liquid) 

and low temperature 

shipping needs 

Ultra-low Temperature Freezers 

Ultra-low temperature freezers 

that combine the highest 

reliability & superior performance 

with cost-effective operation 

and innovative features. 

Cell Culture Media, 

Sera & Reagents 

A complete line of liquid and 

powdered media, process liquids, 

supplements and reagents to 

support cell culture applications 

in academia, research and the 

bioprocessing industry. 

Serological pipettes 

Our Matrix CellMate II, for use 

with graduated and volumetric 

glass and plastic serological 

pipettes, offers simple efficient 

pipetting performance and 

maximum pipetting comfort. 

Cell Culture Vessels 

& Microplates 

A complete line of cellculture 

products and 

bioprocessing systems 

for protein-based drug 

development and 

production. 

Small equipment 

Our range of small 

laboratory equipment 

including homogenisers, 

colony counter, ultra 

sonic bath, peristaltic 

pumps and block heater 

& block supports your 

laboratories requirement. 

Microplate Readers 

Photometry, fluorometry, 

luminometry, TRF or FP, 

dedicated or multimode 

- when it comes to 

microplate reading, we 

have all the answers. 

Flow cytometry 

Partec offers a wide range of 

compact desktop-sized flow 

cytometry systems with up to 

5 light sources and up to 16 

optical parameters, covering 

dedicated applications from 

scientific and industrial routine 

up to high-end research. 

Software 

The flexible and intuitive 

interface of Nautilus 

graphically maps laboratory 

workflows to meet the needs 

of even the most dynamic 

environments 

Cryopreservation Systems 

Cryopreservation systems 

designed to meet the rigorous 

storage requirements of 

cryogenic samples. These 

cryogenic storage systems are 

engineered and built to ensure 

maximum sample protection 

and customer peace of mind. 

Temperature control 

We offer a number of 

professional temperature 

measuring instruments 

to fulfil the individual 

requirements of your 

temperature measurement. 

Antibiotics & Antimycotics 

our antibiotics and 

antimycotics meet rigorous 

quality control tests, 

including verification of 

potency, purity, and solubility 

using microbiological and 

chromatographic methods. 

Successfully cultivating mammalian 

cells requires that you maintain a 

safe, contaminant-free environment 

for your cells and provide optimal 

nutrition for strong, healthy cell 

growth. As your cell lines grow and 

expand, assuring batch-to-batch 

consistency is critical to ensure the 

reproducibility of your experiments, 

regardless of scale. 

Our equipment, consumables 

and media products help ensure 

productive, efficient cell growth and 

passage by providing: 

• Stable growth conditions 

• Outstanding sample protection 

• Optimal nutrition 

Our range includes: 

• Antibiotics and antimycotics 

• Incubators 

• Cell culture media, sera 

and reagents 

• Pipette aids 

• Pipettes 

• Plasticware 

• Shakers 

• Temperature control 

• Water baths 

From key brands: 

• Fisher BioReagents 

• Testo 

• Thermo Scientific HyClone 

• Thermo Scientific Nalgene 

• Thermo Scientific Nunc 

• Thermo Scientific Heraeus 

• Thermo Scientific Matrix 

Pipetting & Liquid Handling 

Handheld and automated 

pipetting systems are 

trusted products that 

include Thermo Scientific 

Finnpipette and Thermo 

Scientific Matrix. We’ve 

expanded our offering with 

additional consumables 

from MBP, including ART 

Aerosol Resistant Tips. 

Water Purification 

We have experts in water 

purification systems who 

can provide information 

and advice on the differring 

grades and volumes of 

purified water required in 

meeting the demands of 

your applications. 

Whether your goal is to explore 

the nature of disease, develop a 

novel drug, or engineer cell lines 

with specific characteristics, 

culture and experimentation often 

involves complex techniques and 

protocols encompassing large 

numbers of samples. Transferring 

minute volumes of cell samples, 

reagents and proteins from flasks to 

microplates is a labor-intensive task 

that demands absolute precision. 

Throughout this stage, care must be 

taken to keep cell lines healthy and 

contaminant-free. We help optimize 

your culture and experimentation 

process by enabling: 

• Precise sample handling 

• Stability and integrity 

• Maximum operator comfort 


• Cell culture plates 

• Cell culture flasks 

• Filtration 

• Laboratory water 

• Cell culture bottles 

• Biological safety cabinets 

• Chamber slides 

• Single channel, multichannel and 

digital pipettes 


• Thermo Scientific Finnpipette 


• Thermo Scientific Max Q 

• Thermo Scientific Forma 



• Thermo Scientific 

• Elga LabWater 

• Whatman Filtration 

Antibodies 

Whether you require 

antibodies for use in 

immunofluorescence 

microscopy, high 

content assays, ELISAs, 

immunoprecipitations, 

Western blotting or many 

other applications, our 

portfolio is now the 

best source for highquality 

antibodies and 

detection reagents. 

Balances 

Equipment and systems 

featuring weighing, 

measurement and 

automation technology 

for laboratory and 

industrial applications. 

Analysing the results of your cell 

culture experiment is the proving 

ground for your innovative ideas. 

Whether determining the results of a 

reporter gene assay, cloning new cell 

lines or staining whole populations 

of cells for characterisation, you 

need efficient, reliable analysis tools 

capable of generating high-quality 

data that validates the accuracy and 

reproducibility of your experiments. 

With the large number of results 

generated, you also need efficient 

sample tracking tools to quickly 

identify the samples and results 

you want to carry forward. We help 

improve the efficiency and quality of 

your analysis by enabling: 

• Streamlined harvesting 

• Efficient, intuitive analysis 

• Seamless data integration 


• HPLC 

• Cell counters 

• Antibodies 

• Homogenisers 

• Laboratory weighing 

• Microplate readers and washers 

• Centrifuges 


• Thermo Scientific 


• Thermo Scientific Pierce 

• Sartorius 

• Partec 

• Santa Cruz 

Cryoware 

Products designed to 

safely contain and organize 

cryogenically preserved 

samples. Protect valuable 

samples safely and 

economically. 

Mr. Frosty 

Provides the critical, 

repeatable -1°C/minute 

cooling rate required 

for successful cell 

cryopreservation and 

recovery 

Cell line and tissue samples 

are precious— in many cases, 

unique—representing a significant 

investment of time and expertise. 

Protecting that investment 

throughout the entire cell culture 

process requires the utmost 

care and precision when storing 

samples and reagents. 

We address this need with 

storage tracking solutions that 

work together to protect valuable 

samples and streamline sample 

retrieval by enabling: 

• Precise temperature control 

• Accurate sample tracking 

• Safe, secure storage 


• ULT freezers 

• Cryogenic preservation tanks 

• Software 

• Cryo tubes 

• Storage racks 

• 2D barcode scanners 

• Tube Picker 

• 2D Cryo tubes and plates 

• Decappers 

• Cryo racks 

• Mr Frosty 


• Thermo Scientific Revco 

• Thermo Scientific ABgene 

• Thermo Scientific Forma 



• Thermo Scientific Matrix 

• Chart MVE

Preventing Cell Culture Contamination with Copper CO 2 Incubators 

Imam El-Danasouri, DVM, Ph.D., HCLD. Director, California Reproductive Laboratories, 1700 California Street #570, 

San Francisco CA 94109 Daniel Schroen, Ph.D. Senior Application Scientist, Thermo Fisher Scientific 


Introduction 

A CO 2 

incubator provides an excellent growth 

environment for cell cultures. However, the same 

warm, humid conditions can also sustain the growth 

of contaminating microorganisms. From easy-toclean 

design to external water reservoirs and heat 

decontamination cycles, Thermo Scientific Heracell® 

CO 2 

incubators are proven to prevent and eliminate 

contamination (1-3) . 

Copper in incubators 

Copper reduces microbes in a wide variety of equipment, including medical 

and scientific devices such as incubators. Years of experience show that 

copper wire, copper sulfate or even pennies added to water reservoirs of CO 2 

incubators significantly inhibit microbial growth. Even better, solid copper 

surfaces clearly reduce the proliferation of contaminants (4) . Thermo Scientific 

Heraeus CO 2 

incubators are available with interiors made from solid copper. 

Because the copper ions do not become airborne, they pose no threat to 

precious cells incubated in culture flasks on copper shelves. 

Antimicrobial action of copper 

Records from early civilizations demonstrate that copper 

can inhibit the growth of many different microorganisms. 

Reviews of modern literature (4) indicate that copper slows 

or stops growth of many organisms, including bacteria, 

fungi, algae and yeast. Copper ions bond to contaminants 

and then disrupt key proteins and processes that are 

critical to microbial life. For instance, the suppression 

of bacterial colonization by solid copper was recently 

demonstrated in Porton Down, England by the Centre 

for Applied Microbiology and Research (CAMR) (5) . In 

that study, CAMR showed that copper piping reduces 

the growth of Legionella pneumophila, causitive agent 

of Legionaire’s Disease. This is the same testing facility 

that certifies many Thermo Scientific centrifuge rotors 

for biocontainment. CAMR also clearly documented that 

the Thermo Scientific Heracell ContraCon heat cycle 

effectively kills fungus and bacteria (2) . 

There are many examples of copper acting as a 

microcide in everyday products, for example: 

• Incorporated into cement or paint, it prevents bacterial 

and fungal growth (for example, in humid basements) 

• It reduces bacteria and algae in cooling systems & towers 

• Copper plumbing pipes reduce the threat from the 

bacteria Legionella pneumophila 

• Brass (copper/zinc alloy) used in machining coolant 

filters removes bacteria and algae 

• Copper-sulfate and -chelate aquacides control aquatic 

pests in ponds and municipal water supplies 

• Copper-based pesticides control nematodes and fungi 

Dr. Imam El-Danasouri, a longtime 

user of copper incubators 

was recently interviewed: 

“In addition to your position 

at California Reproductive 

Laboratories, what other 

positions have you held?” 

Dr. Danasouri “Professor of OB/ 

GYN, Chieti University, Italy, 

Scientific Director of the European 

Institute of Reproductive 

Endocrinology and Infertility, 

Chieti, Italy, and Scientific Director 

at the Institute of Reproductive 

Endocrinology, Ulm, Germany.” 

“Please describe your experience 

with CO 2 

incubators” 

Dr. Danasouri “I have used CO 2 

incubators for research as well as 

for the culture of cells from different 

species for more than 25 years.” 

“What is your experience of using 

Heracell Copper incubators?” 

Dr. Danasouri “I first used copper 

CO 2 

incubators in 1989 at Stanford 

University Medical School. Since 

then, I have been using only copper 

incubators in all the laboratories I 

supervise in the USA, Germany and 

Italy. Recently, I ordered two new 

copper incubators for the Egypt Air 

Hospital in Cairo, Egypt.” 

“Why did you choose copper 

incubators?”Dr. Danasouri 

“Infection within the incubator is 

detrimental to the cells. Since copper 

inhibits bacteria or fungus growth on 

copper surfaces, copper incubators 

reduce the possibility for infection in 

the humidification water or on 

the incubator walls. Studies on the 

contamination of cell culture media 

with heavy metals have shown 

that there are no traces of copper in 

media from the copper incubator.” 

“What types of cells have you 

used with copper incubators?” 

Dr. Danasouri “I have used copper 

incubators for human and mouse 

cultures. I have also cultured many 

other cell types in the copper 

incubators, such as endometrial, 

epithelial and stromal cells, and tubal 

epithelial cells from monkeys, cows 

and humans. Many other cell lines 

have been cultured successfully in 

the incubators.” 

References – (1)Incubators with Thermal Disinfection Cycles (2000). Genetic Engineering News. 20:37. (2)Eliminate Incubator Contamination with Thermo Scientific Heracell. Application Note AN- 

LECO2ELIMCON-1 107 Decontamination Cycles in Heraeus BBD 6220 and Heracell Incubators Completely Eliminate Mycoplasma. Application Note ANLECO2DECONCYC-1 107 (4)Copper Development 

Association, 260 Madison Avenue, New York, NY 10016, 212-251-7200 Ph, 212-251-7234 Fax, Staff@cda.copper.org, www.copper.org (5)The Influence of Plumbing Material, Water Chemistry and 

Temperature on Biofouling of Plumbing Circuits with Particular Reference to the Colonization of Legionella Pneumophila (1993). ICA Project 437B 

19

Feature article 

Stem Cell Promise– 

Research Brings Autograft Revolution Closer 

Stem cells have shown the 

promise to revolutionise the 

treatment of many diseases, 

as noted by George Wolff in his book ‘The Biotech 

Investor’s Bible’: “... The damaged brains of Alzheimer’s 

disease patients may be restored. Severed spinal cords 

may be rejoined. Damaged organs may be rebuilt. Stem 

cells provide hope that this dream will become a reality.” 

Professor Anthony Hollander, the ARC Professor of 

Rheumatology & Tissue Engineering in the Department 

of Cellular & Molecular Medicine at the University of 

Bristol, UK, is in the vanguard of this groundbreaking 

research area. His group has perfected stem cell culture 

protocols that provide the consistent starting material 

essential for all areas of their bioengineering research. 

Furthermore, their knowledge and facilities were 

instrumental in the first ever bioengineered tracheal graft. 

Stem cells show therapeutic potential 

For Professor Hollander and his colleagues, stem cells 

provide the basis for their tissue engineering research. 

Much has been written and discussed on embryonic 

stem cells, but for Professor Hollander’s group, the 

main focus has been on adult (somatic) stem cells. 

Prof Hollander commented, “Embryonic stem cells do 

have the potential to become every type of cell in the 

body, but they are very difficult to control fully – they 

form tumours relatively easily. Somatic stem cells do not 

possess the same breadth of differentiation capabilities 

as embryonic stem cells, but are more predictable and 

controllable.” Importantly, somatic stem cells are found 

in a number of locations, such as the bone marrow, 

and can therefore be retrieved directly from patients. 

This means that it is possible for grafts to be grown from 

these cells and then reimplanted in the same patient – so 

called autologous grafts. This removes the need for 

immunosuppressive therapies to prevent rejection, 

thereby greatly increasing the chance of grafting success. 

Research provides foundation for tissue replacement 

Bone marrow mesenchymal stem cells (BMSCs) 

harvested from the heads of femur bones are the major 

source of stem cells in Professor Hollander’s lab. Dr Sally 

Dickinson, a research associate in the group explained, 

“Bone marrow mesenchymal stem cells are donated 

by patients undergoing hip replacement operations 

and are the perfect starting point for our research, as 

they are multipotent and can therefore form the major 

cell types involved in rheumatology applications.” The 

donated cells are suspended in a specialised stem cell 

culture medium formulated to promote the growth 

and differentiation of BMSCs. To remove any bone 

remnants, the cells are washed several times in the 

medium and fat is then removed by gently centrifuging 

the cells at 1500 RPM for 5 min and recovering the 

cell pellet. Once the cells are clean and free from bone 

or fat, they are seeded in 175 cm2 culture flasks at a 

density of 5-10 million cells. The cells are then placed in 

a CO2 incubator (5% CO2, 37 ºC, 95% Humidity), with 

media changes after four days and then every other day 

until adherent cells have reached 90% confluence. 

Differentiation 

Once successfully expanded, the BMSCs are further 

incubated in specially developed media to enable 

differentiation into either chondrogenic monolayers, 

osteogenic or adipogenic cultures. Alternatively, 

BMSCs can be added to polyglycolic acid (PGA) 

scaffolds and incubated for five weeks with regular 

media changes to create three-dimensional engineered 

cartilage. The majority of research work conducted 

in Prof Hollander’s lab focuses on chondrogenic 

cultures, either monolayer or 3D, as these are the most 

important cell type for osteoarthritis applications. 

Analysis 

Several analytical techniques are used to assess BMSC 

cultures and their differentiation, including histological 

staining and real-time PCR. However, the bulk of the 

analyses on the engineered cartilage are carried out using 

enzyme-linked immunosorbant assays (ELISAs), since 

they provide quantitative biochemical measurements 

for key molecules such as collagen types I and II. All 

ELISAs in Professor Hollander’s lab are analysed on 

a photometer.A range of laboratory instruments are 

used in the research, many of which are from the 

Thermo Scientific Stem Cell Excellence portfolio.* 

*Instruments used from this range include the Thermo Scientific Sorvall Legend RT-Plus centrifuge, the Thermo Scientific Cytoperm 2 CO 2 

Incubator, and the Thermo Scientific 

Multiskan microplate reader. At every manipulation stage, the cells were handled within a Thermo Scientific Herasafe KS12 Type 2 Class 2 biological safety cabinet. 

Cell culture research also requires a large amount of manual pipetting which – if not done properly – can lead to inconsistencies and possibly repetitive strain injuries for 

the user. In Prof Hollander’s lab, the group used Thermo Scientific Finnpipettes. Essential reagents, which contribute to the ongoing success of the lab, were stored in a 

Thermo Scientific Revco freezer, which provides ultra low temperature storage at -86ºC. 

20

Clinical collaborations lead to patient therapies 

As one of the foremost scientists in the rheumatology 

field and the development of chondrogenic cultures from 

BMSCs, Professor Hollander works closely with clinical 

teams to develop patient-specific cartilage autografts. 

These are generated by extracting cartilage cells (rather 

than BMSCs) from the patient and culturing them to 

provide autologous chondrocytes, which are then seeded 

into a three- dimensional biodegradable material (derived 

from the total esterification of hyaluronan with benzyl 

alcohol and constructed into a non-woven configuration). 

These engineered grafts are then placed at the site of 

the cartilage injury, often without the need to glue or 

suture them in place. Furthermore, the procedure does 

not necessitate open surgery since a miniarthrotomy 

is usually sufficient. Once in place, the graft quickly 

integrates with the patient’s existing tissues, providing 

good collagen composition and integration with the 

underlying bone. The autograft technique provides 

several distinct advantages, namely less stressful surgical 

procedures and perhaps more importantly, the lack of 

any immune response. This is a major advance over 

allografts, which require the use of powerful immunesuppressing 

drugs for extended periods post-transplant. 

The first ever bio-engineered tracheal graft 

Last summer, Professor Hollander received a request 

for help from a friend and colleague, Professor Martin 

Birchall, a surgical professor at the University of Bristol. 

A patient of Dr Birchall’s had suffered serious damage to 

her trachea as a result of contracting tuberculosis (TB). 

The patient, was a young mother whose only chance 

of survival at the time was to have one lung removed, 

which would have seriously affected her quality of life 

and her ability to look after her children. After much 

discussion, Professor Hollander and his team very 

quickly set to work adapting their existing osteoarthritis 

based protocols to enable Professor Birchall to grow 

a large population of chondrocytes derived from the 

patient’s BMSCs. A section of human trachea was 

donated for use as a scaffold on which the new tissue 

could be grown. The trachea was stripped of the donor’s 

cells, leaving a trunk of non- immunogenic connective 

tissue onto which the chondrocytes were seeded. This 

seeding process used a novel bioreactor developed 

at the Politechnico di Milano, Italy, which provided the 

right environment for the cells to form the cartilaginous 

part of the trachea within four days of seeding. The graft 

was then lined with epithelial cells and transplanted 

into the patient, who responded very quickly to the 

new airway section, without any sign of rejection (no 

antibodies to the graft were found). Subsequent biopsies 

have shown that the new section is fully integrated 

with the existing airway and is fully supplied with blood 

vessels. She is now able to live life as if she had not 

been struck down with TB, a result that would never 

have been possible if her lung had been removed. 

Discussion 

Stem cell based-therapies have promised huge changes 

in the treatments of many diseases and disorders, but 

much research is still required to ensure safety and 

consistency before they can be applied more extensively. 

Prof Hollander and his colleagues at the Department 

of Cellular & Molecular Medicine at the University 

of Bristol have been investigating the fundamental 

principles governing the differentiation of bone marrow 

stem cells into chondrocytes – the source of cartilage. 

Through this research they aim to further improve 

the processes used to generate chondrocyte-based 

autografts, which have already started to prove their 

value in the treatment of cartilage damage. Throughout 

their pioneering research, Professor Hollander’s team 

has come to rely on the dependability and functionality 

of a broad array of standard and advanced laboratory 

equipment specifically designed to provide the highest 

quality and reliability in the cell biology laboratory. As 

a result of their dedicated work, Professor Hollander’s 

team was able to take part in the amazing feat of the 

first ever bio-engineered tracheal graft. Their work 

has enabled this patient to regain an amazing quality 

of life following a life-threatening condition while 

increasing the drive among researchers and clinicians 

to more expansive use of stem cell based therapies. 

Feature article 

Professor Anthony 

Hollander, ARC 

Professor of 

Rheumatology & Tissue 

Engineering in the 

Department of Cellular 

& Molecular Medicine 

at the University of 

Bristol, UK. (Image 

courtesy of Dr Sally 

Dickinson, University 

of Bristol). 

Figure 1. Adherent 

human adult bone 

marrow stem cells in 

culture. (Image courtesy 

of Dr Sally Dickinson, 

University of Bristol) 

Figure 2. Tissue 

engineered cartilage 

produced from 

bone marrow stem 

cells (Macroscopic 

Appearance). (Images 

courtesy of Dr Sally 

Dickinson, University 

of Bristol). 

21

Article 

Choosing the Right Centrifuge for Your Application 

Ms. Goodman – Sample Preparation and Separations Applications Product Manager, Thermo Fisher Scientific Inc 

Figure 1–Superspeed centrifuges are an excellent choice for multiuser, multiprotocol environments, offering high capacity, high g-forces, and a broad range of rotors and accessories. 

With so many recent advances in both science and 

technology, it is wise to educate yourself about the wide 

range of centrifuge options now available. Following are 

some key questions that can be used to determine the 

type of centrifuge that will best meet your needs: 

1. What applications and protocols will the centrifuge 

be used to support? 

2. What are the maximum and minimum g-force 

(relative centrifugal force, RCF) and volume 

requirements? 

3. How many tubes or samples must be processed in a 

run, shift, or day? 

4. What types of sample formats will the centrifuge 

need to support (i.e., microplates, blood collection 

tubes, disposable conical tubes)? 

5. What type of rotors will be needed to support your 

applications (i.e., fixed angle, swinging bucket)? 

6. How many people will be using the centrifuge? 

7. Is versatility important? That is, do you anticipate 

the need for a broad range of protocols and 

multiple users, or will you be performing the same 

standardized protocol day after day? 

8. Do you have any space restrictions, such as benchtop 

or floor space only? 

9. Do you have special needs such as process 

traceability, user lock-out, or biocontainment? 

10. What is your budget? 

Once you have answered these questions, you should 

have a better picture of your centrifuge requirements. It 

is also helpful to review the basic types of centrifuges to 

make sure you know which category best fits your needs. 

Floor model or benchtop? 

Centrifuges are generally classified 

as either floor-standing or benchtop 

models. The style you choose is 

typically determined by performance 

requirements, available space, and 

budget.Floor-model centrifuges 

free up bench space and are often 

chosen for either high-speed or 

high-capacity sample processing. 

Within the floor-model category 

there are superspeed centrifuges, 

ultracentrifuges, and low-speed 

centrifuges.Benchtop centrifuges 

offer versatility and convenience, and 

can be equipped to accommodate a 

broad range of needs, making them 

a cost-effective solution for many 

laboratories. Benchtop platforms 

include general-purpose centrifuges, 

microcentrifuges, small clinical 

centrifuges, cell washers, and 

high-speed models. 

Choosing the right floormodel 

centrifuge 

Superspeed centrifuges – If you 

are looking for high capacity, high 

g-force and versatility, a superspeed 

centrifuge (Figure 1) is probably 

the best choice. Many superspeed 

models offer a choice of up to 40 

rotors, making them an excellent 

solution for core laboratories 

performing general preparative 

applications such as whole cell 

separations, protein precipitation, 

tissue culture, subcellular isolation 

(i.e., Golgi bodies, ribosomes), 

plasmid preps, and DNA/RNA 

separations. Superspeed centrifuges 

are also the best option for multiuser, 

multiprotocol environments. Their 

versatility enables researchers 

to step into new, cutting-edge 

technologies without purchasing a 

dedicated centrifuge for one specific 

application. 

Ultracentrifuges – If your 

application calls for g-forces of up 

to 1,000,000 × g, you will need an 

ultracentrifuge. These extremely 

powerful centrifuges support sample 

volumes up to 250mL. Within the 

ultracentrifuge line there are two 

platforms: full-size floor models, 

which support g-forces of up to 

802,000 × g and volumes up to 

250mL; and micro-ultracentrifuges, 

which support g-forces of above 

1,000,000 × g and microvolume 

samples up to 13.5mL. 

Common ultracentrifuge applications 

include the separation of virus 

particles; DNA, protein, or RNA 

fractionation; as well as lipoprotein 

flotation. Density and size gradient 

22

Article 

separations are also regularly 

performed in ultracentrifuges and, 

more recently, a number of new 

nanotechnology applications have 

appeared. 

Low-speed centrifuges – These 

large-capacity centrifuges have 

fairly specific applications due to 

the maximum RCF of approximately 

7000 × g. The most common lowspeed 

application is the separation of 

whole cells, for example, separating 

red blood cells and platelets from 

whole blood. The second most 

common application is the whole 

cell harvest step in the processing 

of large volumes of cultures from 

bioreactors in the bioprocessing and 

pharmaceutical industries. Lowspeed 

centrifuges offer a basic set of 

rotors that support volumes ranging 

from 1.5mL (with adapters) up to 

2000mL. 

Choosing the right benchtop 

centrifuge 

Benchtop centrifuges are 

available in a wide variety of 

platforms designed for different 

application requirements, including 

general-purpose benchtops, 

microcentrifuges, small clinical 

centrifuges, cell washers, and 

high-speed benchtop centrifuges. 

General-purpose benchtop 

centrifuges – These workhorse 

units (Figure 2) are the most 

common type of centrifuge found 

in the laboratory. Their versatility 

makes them extremely practical: 

They offer a wide range of rotor 

types, volumes, and speeds within 

a single unit to meet the demands of 

many common protocols. Generalpurpose 

centrifuges are typically 

used for tissue culture, DNA/RNA 

research, cell harvesting, subcellular 

separations, protein work, and many 

other applications. Most generalpurpose 

units can be equipped with 

a broad range of swinging-bucket 

and fixed-angle rotors, making 

them an excellent fit in a multiuser 

environment where floor space is an 

issue and RCFs of up to 24,000 × g 

are sufficient. 

Microcentrifuges – Like the 

general-purpose benchtop models, 

microcentrifuges (Figure 3) are a 

necessity in every laboratory. These 

compact units provide RCFs of up 

to 21,000 × g, which is sufficient 

for the most common microvolume 

applications such as plasmid, DNA/ 

RNA work, and mini-prep kits. 

Designed to spin up to 2mL volumes, 

microcentrifuges are typically 

equipped with rotors that accept 

commonly used 0.2-mL PCR tubes 

and 1 .5-mL/2.0-mL disposable 

micro- centrifuge tubes and filters. 

Clinical benchtop centrifuges – 

These compact models are designed 

for use in hospitals and clinics that 

require a low-throughput unit to 

spin blood collection tubes and 

urine samples at very low speeds 

for diagnostic examination. Most 

of these centrifuges spin at RCFs 

at or below 3000 × g. The volume 

supported by a typical clinical 

benchtop unit ranges from 3-mL up 

to 15-mL tubes, with throughput 

ranging from 4 to 28 tubes per run, 

depending on the tube size and unit 

selected. 

Cell washers – These specialpurpose 

centrifuges support very 

specific applications in the clinical 

and medical industry: washing away 

cellular debris, extraneous proteins, 

and other constituents of donor 

blood from red blood cells. 

The washed red blood cells are 

used for tests such as crossmatching 

prior to blood transfusion. 

Cell washers spin at RCFs at or 

below 1500 × g and support 3-mL 

and 5-mL culture tubes. 

High-speed benchtop centrifuges 

These powerful and compact units 

offer g-forces close to that of a floormodel 

superspeed centrifuge, but 

with a limited set of rotors. The most 

common rotors and applications for this type of centrifuge 

include a low-speed, high-volume swing-out rotor for a 

whole cell harvest step; a high-speed, midvolume (i.e., 

50–15 mL) fixed- angle rotor for a subcellular pelleting 

step; or a high-speed, low-volume (1.5-mL/2.0-mL) fixedangle 

rotor for certain DNA/RNA applications. The average 

maximum RCF for a high-speed benchtop is 50,000 × g, 

with a volume range from 1.5 mL to 200 mL over a range of 

swinging-bucket and fixed-angle rotors. 

Understanding the different types of centrifuge platforms 

and the applications they support is a good first step in 

the selection process. Once you have identified your 

application needs and the appropriate centrifuge model, 

your centrifuge supplier should be able to assist you 

in configuring the best system (centrifuge, rotor, and 

consumables) to meet your specific requirements. 

For further information please contact: Dhru Patel (AU) 

dhru.patel@thermofisher.com or Ph: +613 9757 4522 

Kris Baker (NZ) kris.baker@thermofisher.com 

Ph: +64 9 980 6763 

Figure 2 –General-purpose benchtop centrifuges offer versatility and convenience, 

and can be equipped many ways to support tissue culture, DNA/RNA research, cell 

harvesting, subcellular separations, and many other applications. 

Figure 3 –Microcentrifuges are a necessity in every laboratory for everyday microvolume 

applications such as plasmid, DNA and RNA work, and miniprep kits. 

23

Latest news 

The next generation in Automated liquid handling 

The new Thermo Scientific 

Versette automated 

liquid handler 

Compatible with 19 interchangeable, 

RFID-tagged pipetting heads, the 

Versette provides exceptional 

pipetting versatility for a broad range 

of applications that require single to 

384 channel automated pipetting. 

To further optimize performance, 

the single, 8 and 12-channel pipetting 

heads utilize the Thermo Scientific 

ClipTips, which securely seal to 

the pipette head. The unique clip 

design requires minimal insertion 

and ejection force, decreasing wear 

and tear and increasing the life of the 

instrument and pipetting heads. 

The 96 and 384 channel pipetting 

heads use the Thermo Scientific 

D.A.R.T.s tips, which seal to ensure 

accurate and precise pipetting 

across all channels. 

With a compact footprint, further facilitated by the unique 

dual-level design of the 6 position stage, the Versette can 

be placed exactly where it is needed, making it ideal for 

use on any benchtop or in an enclosure. 

Simple and complex pipetting procedures can be 

programmed quickly and easily via the intuitive onboard 

LCD user interface or the Thermo Scientific ControlMate 

software. This advanced instrument control, combined 

with precision and versatility, makes the Thermo 

Scientific Versette automated liquid handling platform 

ideal whether transitioning from handheld pipetting or 

establishing an integrated liquid handling system. 

For more information visit 

www.thermoscientific.com/versette or call 

Mika Mitropoulos (AU) on 9757 4474 

Mika.Mitropoulos@thermofisher.com 

Kris Baker (NZ) +64 9 980 6763 

kris.baker@thermofisher.com 

The Versette platform is packed 

full of features, it offers two stage 

capacity options. These easy to 

swap 2 and 6 position stages provide 

flexibility for both stand-alone and 

robot-friendly use. 

24

When is a μL not a μL? 

Have you ever picked up your pipette and thought… Is this accurate? 

Latest news News 

Pipettes are used to measure 

and transfer liquids; a simple 

enough processes until you 

really think about it. 

What if it isn’t reliably aspirating 

the correct amount of liquid? 

What will this do to your results? 

Can you rely on the results you 

are getting if your pipette is not 

calibrated? The answer is…no. 

The pipette is one of the most commonly used instruments in labs everywhere. Many pipettes are 

shared; they are used with varying liquids and are treated differently by different users. Pipettes 

work by creating an air gap to aspirate and then dispense liquid. The air gap is proportional to the 

amount of liquid that is being dispensed and it can be affected by many different factors. 

Usage over time, damage and general wear can have a major impact on dispensing accuracy and 

precision. If your pipette has not been checked and calibrated, the quality of the results obtained 

is questionable. It may not seem like a major factor at the time, but repeated incorrect dispensing 

can have a cumulative impact on results and can lead to major errors or even test failures. 

Next time you use a pipette, think about the quality of the results you want to achieve and whether 

the pipette in your hand is going to deliver. 

Imagine the peace of mind that comes from knowing that all your valuable 

samples are being dispensed reliably every time you pick up your pipette. At 

Thermo Fisher Scientific we know that selecting the right pipette calibration 

service is important for the success and efficiency of your laboratory. We also 

know that each lab can vary considerably and that’s why we have tailored our 

pipette service centre to meet your diverse needs. 

We let you choose the services you want 

Choose between 3 different modules for your pipettes: 

Benefits of our NATA accredited 

pipette calibration centre are: 

• Nata certified laboratory + 

equipment 

• Accredited for all pipette 

calibrations from volumes 

0.2µL to 60mL 

• 5 and 6 place balances as per 

AS2162.2-1998 

• Speedy turnaround times 

• Loan pipettes (contract 

customers) 

• We are the manufacturer of the 

Finnpipettes, therefore easier 

access to replacement pipettes/ 

warranty claims plus original and 

cost effective spare parts 

• Relationships with all the other 

pipette manufacturers with 

access to their spare parts and 

warranties 

For further information email : 

ServiceAU@thermofisher.com 

1. Maintenance & Calibration 

• Internal & external inspection, cleaning 

and lubrication 

• Replacement of seal/o-ring 

if necessary 

• NATA calibration certificate 

• Calibration sticker affixed to pipette 

to provide a suggested date for next 

calibration/service 

2. Training & Education 

• Over our many years of experience, we 

have developed first rate user training 

methods to ensure your staff correctly 

use and maintain their pipettes 

• Our training workshops include correct 

pipetting techniques, maintenance, 

servicing and calibration of pipettes 

• Workshops also cover AS/ISO standards, 

equipment recommendations and 

general requirements for setting up 

in-house calibration services 

• Onsite or depot training options 

are available 

WARRANT Y 

EXTENSION 

CONTAINED 

SERVICE COSTS 

MAINTENANCE & 

CALIBRATION 

INSTALLATION 

INSTALLATION 

REPAIR & 

INSPEC TION 

EMERGENCY 

RESPONSE 

TRAINING 

3. Repair & Inspection 

• Internal & external inspection, cleaning and lubrication 

• Replacement of seal/o-ring if necessary 

• Replacement of worn/broken parts 

• Free of charge repair evaluation – economical or 

uneconomical to repair? 

• NATA calibration certificate 

• Calibration sticker affixed to pipette to provide a 

suggested date for next calibration/service 

25

Latest news 

Chart MVE for Ultra Cold Storage 

Thermo Fisher Scientific is proud to announce it is now the distributor of choice 

for Chart MVE vacuum insulated products and liquid nitrogen freezers. 

Excellence & Innovation 

Chart-MVE is the world’s leading manufacturer of 

vacuum insulated products and cryogenic systems. 

More than forty years ago, we set the standard for 

storage of biological materials at low temperatures. 

Today, we continue to exceed these standards. 

Industries from around the world look to Chart-MVE 

for excellence and innovation. Our solutions empower 

industries to better utilize cryogenic technology. In 

this manner, Chart-MVE continues to make a vital 

contribution in today’s biomedical industry. 

Chart-MVE has the solution for all of your cryogenic 

storage needs. We offer the broadest range of storage 

capacities for your biological products, with the most 

advanced vacuum technology available today. 

Chart-MVE is the market leader in the manufacturing 

of Bulk Storage, Liquid Cylinder and Vacuum Insulated 

Pipe products. Chart-MVE applied this knowledge 

to the development and creation of “Turn Key” liquid 

nitrogen supply systems that can provide your freezer 

with the most economical use of liquid nitrogen and 

the best return on your storage investment. 

Every Chart-MVE freezer is designed for optimum 

vacuum performance for the duration of its use. 

Chart-MVE freezers are engineered to hold and 

maintain specific temperatures, whether samples 

are in liquid or vapour. 

Chart-MVE offers the widest range of storage capacities 

and storage options (from -125°C to -196°C) to suit your 

biological product needs. By choosing Chart-MVE, you 

are installing a secure and viable environment, free of 

the noise and heat created by mechanical refrigeration 

systems. 

Chart-MVE products meet worldwide standards of 

excellence such as CE, MDD, UL, IATA, TGA, and ISO 

9001. Factory tested to ensure reliability in the field, 

Chart-MVE vessels are backed by one of the strongest 

and longest warranties in the industry. 

Product Warranties 

• Standard two (2) year warranty on all equipment. 

• Three (3) year vacuum warranty on CryoSystem 

Series, Doble Series and Vapor Shippers. 

• Five (5) year vacuum warranty on Stainless Steel 

Freezers, XC/SC Aluminum Units, and LAB units. 

• Static evaporation rate and static holding times 

are nominal. Actual rate and holding time will be 

affected by the nature of container use, atmospheric 

conditions, and manufacturing tolerances. 

For further information please contact: David Felici (AU) 

david.felici@thermofisher.com or Ph: +613 9757 4396 

Kris Baker (NZ) kris.baker@thermofisher.com or 

Ph: +64 9 980 6763 

26

Culture serum Q & A 

Latest news 

How should I condition my culture 

to new sera & media? 

For best results, the process of adapting cells to a novel 

culturing milieu should be sequential. Direct migration to 

a new serum or media can be detrimental to cell viability 

and should be avoided. The most common procedure is 

to dilute the culture in a low percentage of new media at 

the time of passage, increasing gradually in proportion 

upon each subculture (split). 

What are the benefits of using 

animal-free media? 

Animal-free media contain no 

animal-derived components and 

have defined components which 

eliminate any inconsistencies 

associated with serum batch 

variations. Hence, it is not necessary 

to conduct time consuming 

lot-testing of serum supplements 

in order to acquire reliable and 

reproducable results. Media free of 

animal components are commonly 

used in the production of human 

and animal biopharmaceuticals, 

diagnostic reagents and 

bioagricultural products. It is also 

beneficial for cell culture applications 

aimed at high-throughput production 

of recombinant proteins, monoclonal 

antibodies and viral vectors, as 

reduced protein content improves 

both purification yield and efficiency. 

Do I need heat inactivated sera 

for general cell cultures? 

The heat inactivation procedure 

involves warming thawed FBS to 

between 45°C to 62°C for 15 to 60 

minutes, and was once regularly 

performed to destroy components 

of the complement cascade and 

neutralise mycoplasma in the sera. 

However, recent findings suggest 

that this practice is often not 

essential for many cell culture 

applications and may instead 

negatively affect growth rates as 

vital temperature-sensitive nutrients 

are also destroyed in the heating 

process. Vigorous sterile filtration 

of HyClone FBS through three 

consecutive 0.1µm pore-size rated 

filters ensures only minuscule level 

of complement and mycoplasma 

persists in the serum. 

To have you questions answered email us at: Bio-Innovation@thermofisher.com 

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27

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