PELONews April/May

pelobiotech

PELONEWS

WITH FOCUS ON PRIMARY CELLS

PELOBiotech Newsletter April /May 2019

“Science without religion

is lame, religion without

science is blind.“

-Albert Einstein, Aus meinen späten

Jahren, Selbstporträt, 1936) -

Dear Scientist,

just a few more days and then it's finally Easter. We also have

some neat Easter surprises for you. Have a look at our offers

at www. pelobiotech.com. Happy Easter to you all.

Christiane Büchsel Editor in Chief PELONews

New video about Drug Development platform: CellPrism ®

...is a drug development platform offering customizable cell-based in vitro

assays using optimally sourced and processed primary cells (blood and

bone marrow cells) from multiple species.

-> Learn & read more here or Watch & Learn via video.

Content

Let‘s Talk: Learn from the best

about 3D Models

CSO-Talk about

Primary Cells

White paper: Off Target Toxicity&Linker

Stability

New Products: Ultralow Cell

Binding Cultureware & Fluorescent

Dyes

Paper Alert: NEW Hydrogel

...like a natural muscle

Featured Fields

&Products

Play






Immuno-Oncology

Primary Cells

3D models

Cytotoxicity

ADCs


PELONews

We have talked: Optimised Cancer

Treatments and EV research with 3D Models

To an expert- talk about “Optimised Cancer Treatments and EV research with 3D Models”

invited PELOBIotech GmbH and its Swiss partner abc biopply at the IZB Sky-Lounge end of

March 2019. Around 20 scientists from Bavarian biotech companies and institutes such as

TUM, LMU, Helmholtz Centre in Munich and Fraunhofer in Regensburg discussed with highlevel

cancer research experts Dr. med. Andreas Thomsen and Dr. Irina Nazarenko from Universitätsklinikum

Freiburg about new models for tissue-specific interaction of cells and how

they optimize their treatments.

First Dr. Marco Leu, Co-Owner abc biopply, gave a short overview of a novel and innovative 3D microwell array in

the analysis of adhesion independent micro-organoids, named 3D CoSeedis. “I am glad abc biopply was able to

collaborate with Dr. Andreas Thomsen and to further develop this unique and novel technology. We are proud

to have recently released the first standardizes systems it to the scientific world” says Dr. Leu. 3DCoSeedis allows

the support of the growth of feeder cells in the formation of 3D cell constructs that are not in direct contact with

the test cells themselves.

„It’s totally cool!“

Both key speakers are already working with the new system 3D CoSeedis, as in fact Dr.

Thomsen invented the system himself. His lecture “Creating Unique Microenvironments

in 3D Models” showed a broad range of applications and inspired the researchers on their

projects on tissue-specific interaction of cells. A lot of wows he earned showing his results

in comparison with standard 2d methods and the new 3D model. “It makes it so much easier”,

the Freiburg radio-oncologist says. His survey “Do you enjoy colony formation assays?” performed among

colleagues made the audience laugh. They knew almost everybody dislike it and now are given a possibility to

work much more efficient.

A lot of interest had the lecture by Dr. Irina Nazarenko: Studying Intercellular Communication by the mean of

Extracellular Vesicles in 3D Models. She talked about the impact environment on production, cargo, and function

of extracellular vesicles (EVs) in several cancer models. EVs mediate intercellular communication between tumor

and tumor stroma, tumor and immune system, playing a pivotal role in metastases, immunosuppression, angiogenesis

and other systemic effects in progressing tumor. In her experience 3DCoSeedis TM is applicable for EV isolation,

analytic and functional characterization. Using the 3D microwell array more EVs/cell can be isolated in a

considerably more cost- and efforts- efficient manner than 2D culture. “My co-workers and me love working with

it”, Dr. Nazarenko says, as the cells are growing in more physiological environment and producing more EVs. 3D

environments are likely to trigger release of smaller EVs with increased amounts of certain miRNAs and decreased

amounts of their target proteins. She also emphasized that functional impact of 3D environment on EVs

differs between cell lines and should be individually analyzed.

„We are already switching!“

Dr. Nazarenko is more than convinced by the 3D CoSeedis system. Asked whether she would

switch if she had the opportunity: “Definitely, it´s genius, totally cool. So, where it makes

sense, we are already switching, step by step”, she outlines. One of the participants confirms

her point of view: “We are more and more searching for 3D models, also in EV research. So

this system sounds very promising to us.”

Peek into presentation Dr. Irina Narzarenko and download paper here.

Download presentation Dr. med. Andreas Thomsen, and download paper here:

Peek into presentation Dr. Marco Leu


Here some

impressions of

this great

3D model

symposium end

of March.


PELONews

Primary Cells

CSO-Talk: Why Isolation and Cultivation of PCs is not always easy

by Dr. Lothar Steeb

Primary cell culture is the only basis for developing alternative test systems and methods. Theoretically, if one

reads the method parts of publications, primary cells can be obtained and cultured quite easily.

However, anyone who has already dealt with this topic, the isolation and cultivation of primary cells from human

tissues, will quickly notice that there are some challenges that are not mentioned in every methods and that these

cells can only show their advantages if they are cultivated differently from permanent cell lines with special microenvironment

conditions. An endothelial cell does not grow in an environment for neurons or epithelial cells and

vice versa. In addition, the processing of donor tissue must always be based on infectious material, which requires

stringent methodological work. Nevertheless, the advantages of primary cells are so great that it is worth working

with them.

Primary cells are of particular importance with regard to

- metabolism

- morphology

- physiology

- Genetic characteristics

- demeanor.

The most in vivo state is the one closest to the body and therefore form the link

CSO: Dr Lothar Steeb

between the body and ex vivo cultures. Cross-contaminations and false characterizations can also be excluded.

The markers are usually unambiguous. Another disadvantage of cell lines is that they often differ genetically and

phenotypically from their original tissue. In contrast, primary cells retain many of the important markers and functions

observed in vivo. For example, endothelial cell lines lack various functional markers, while primary endothelial

cells retain these important characteristics. In animal cells, in particular, an advantage is extremely important

because it enables cross-generational and genetically modified investigations.

Many years ago, PELOBiotech was one of the first suppliers of cells that recognised the increasing demands on cell

culture media and the cells‘ environment represent an opportunity not only to maintain the individuality of cells,

but also to increase the comparability of the results. By creating defined and standardised environmental conditions

the results from different working groups are more stringent and also more reliable. They cannot be influenced

by unknown factors such as serum components or raw products such as matrices or extracts.

With its wide range of products, PELOBiotech offers exactly these opportunities. We are happy to help find the cells you are

looking for (there are currently more than a thousand possibilities open to you), the media selection and all the tools you need

for optimal primary cell cultivation. PELOBiotech stands for solutions and assistance for all questions regarding primary cell

culture and supports in choosing the most efficient, cost-effective and optimized solution from our almost 30,000 products.

Want to know more, click on the following links:

Animal Cells, Media & Tools:

Human cells,Media & Tools


Assays&Arrays

WHITE PAPER | Off-Target Toxicity and Linker Stability

By Emer Clarke, CSO ReachBio Research Labs

CSO Dr. Emer Clarke, PhD

The Primary Cell Experts

ReachBio Research Labs’ scientists use human primary cells

every day in our contract services lab for high value projects for

clients in the pharma and biotechnology industries. Their extensive

experience in handling and using human primary cells is

used in the production of our human cell products, resulting in

high quality cells that you can rely on for your own important

experiments. We use Cryostor CS5 serum-free cryopreservation

medium to ensure high post-thaw viability and overfill each vial

to ensure you always get at least the number of cells we advertise

on the vial when they are thawed correctly.

-> Ask us at PELOBiotech for more.

F

Antibody drug conjugates (ADCs) combine the benefits of both therapeutic monoclonal antibodies (mAb) and potent

small molecule cytotoxic drugs. These moieties are connected through linkers that should be stable within

systemic circulation, while maintaining the ability to cleave within the mAb-targeted cells to release highly specific

payloads.

In addition to the targeted killing of antigen-positive tumor cells, some ADCs appear to have significant off-target

effects, causing neutropenia and thrombocytopenia, which have necessitated some clinical trials being terminated

early. However, the success of some ADCs in treating diseases has stimulated the continued research of these

constructs.

ReachBio has two distinct assay platforms to assess cytotoxicity caused by ADCs

In the initial platform, they can evaluate off-target toxicity using colony forming cell (CFC) assays to assess effects

on erythroid, myeloid, or megakaryocyte progenitors using marrow derived from human, NHP, rat, and mouse.

Our second platform addresses linker stability. They have adopted the neutrophil differentiation assay described

by Zhao et al (2017), whereby neutrophils cause their own demise by releasing enzymes which cleave the payload

from the antibody and thus kill developing neutrophils.

Download Emer`s White paper here


PELONews

New Products

Ultralow Cell Binding Cultureware

HydroCell is a low cell biding cultureware. On its surface, super hydrophilic

polymer was immobilized at nano-thickness. This hydrophilic character helps

the formation of embryoid bodies of ES/iPS cell or Spheroid in culture.: Compared

to other brands' low cell binding cultureware, HydroCell shows lower

cell binding.


• Super hydrophilic polymer is immobilized to the surface of the cultureware

at nano-thickness.

• Lowest attachment rate among similar competitive brands.

• Three formats : Microplates, Dishes and Flask types.

Fig.1 Overview of HydroCell surface


• Formation of embryoid bodies of ES cells in culture

• Formation of anchorage-independent colonies

• Differentiation such as cartilage (spheroid formation)

• Culture & storage of macrophage/immune cells

• Screening of anti-cancer drugs (replacement for soft agar assays)


Fig.2 Comparison to other brand products

Mouse macrophages were incubated for 3 days in (1) HydroCell, (2) another brand's product and (3) non-treated cultureware. Only HydroCell allowed

uniform suspension culture (>99% of the cells remained in suspension), simple cell dissociation and harvesting by pipetting.

For 3D

Cell Culture

Fig. Stem cell sphere formation iPS spheres are formed on HydroCell (left) or other brand low cell binding dish.A complete sphere was surely formed on

HydroCell, while adhesion was observed on other low cell binding dish. (Data provided by A. Umezawa, National Center for Child Health and Development)

For more infos and prices, just call us: 0049 89 517286 59–0


Reversible Fluorescent Dye for Nucleus

NucleoSeeing is a novel live cell imaging probe which emits green fluorescence

by binding to DNA specifically. NucleoSeeing can be used for not only in animal cells or tissues, but

also Guard cell of Arabidopsis thaliana with high S/N ratio. Moreover, NucleoSeeing can be used as a pH sensor in

nucleus. This product has been commercialized under the license from Nagoya Institute of Technology.


NucleoSeeing is composed of green fluorescent dye and DNA binding tag. Under DNA free condition, Nucleoseeing

is in folded conformation and no fluorescent is emitted. However, once it binds to DNA, the conformation

will change andemitts green fluorescence.

Ex/

Em

Fluoresce

nce

Phototoxicity

Nucleu

s

Spe

cifici

ty

Cell

Me

m.

Permea

bilit

y

Cyto-

toxi-

city

Live Fixed

NucleoS

Green Low Yes Yes Low Yes Yes

488/

520

eeing

Hoech 350/

st 461

DAPI 350/

461

Company

X

Company

Y

485/

498

646/

680

Blue High Yes Yes

Hig

h

Yes Yes

Blue High Yes No N/A No Yes

Green Low No Yes N/A Yes Yes

Red Low Yes Yes

Hig

h

Yes Yes


Comparison table of dyes for nucleus

Fig.1 Staining of nucleus in various cultured cells

References

1. Nakamura, A., et al., Chem. Commun., 50, 6149-6152 (2014)

Hoechst tagging: a modular strategy to design synthetic fluorescent

probes for live-cell nucleus imaging.

2.

2. Ueda, M., et al., ACS Cent. Sci., 3 (5), 462-472 (2017)

Noncanonical function of a small-molecular virulence factor coronatine

against plant immunity: an in vivo raman imaging approach.

3 papers is equal to NucleoSeeing.

3. Nakamura, A. and Tsukiji, S., Bioorg. Med. Chem. Lett., 27 (14),

3127-3130 (2017)Ratiometric fluorescence imaging of nuclear pH in

living cells using hoechst-tagged fluorescein. * hoeAc2FL in above

More Product Information/Datasheet here.


PELONews

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Paper Alert from our partner Noviocell

New hydrogel stiffens and

softens like a natural muscle

„A new synthetic hydrogel that becomes up to 50 times stiffer upon heating

just a few degrees has been developed by researchers at Radboud University

in the Netherlands. The stiffening process is reversible, and the team believes

that the hydrogel could be used in a range of new applications including tissue

engineering. The controlled stiffening or softening of materials is very common

in biology, playing roles in processes such as muscle contraction, tissue

fibrosis, the enzymatic degradation of tissues and tumour formation. It involves

a biological cell converting chemical energy into mechanical stresses,

which cause the cell’s cytoskeleton to stiffen with stress. However, mimicking

this ability to stiffen and soften in a synthetic material such as a hydrogel has

proven very tricky to achieve.

Read more here.

We like your feedback – tell us what you love, don’t like so much and what you would

like to get, please. Just reply Please update your subscription anytime, as we like to

comply to the new GDPR guidelines.

How to

reach us

If you need any further

assistance or if you like

what you see, tell us:

PELOBiotech GmbH

Klopferspitz 19 82152

Planegg | Germany

Tel.: +49 89 517286

59 0

info@pelobiotech.com

| www.pelobiotech.com

Managing Directors:

Dr. Peter Frost,

Dr Lothar Steeb

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