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) -
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.
Let‘s Talk: Learn from the best
about 3D Models
White paper: Off Target Toxicity&Linker
New Products: Ultralow Cell
Binding Cultureware & Fluorescent
Paper Alert: NEW Hydrogel
...like a natural muscle
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
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
Primary cells are of particular importance with regard to
- Genetic characteristics
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
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.
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
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
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
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
• Super hydrophilic polymer is immobilized to the surface of the cultureware
• 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.
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.
Green Low Yes Yes Low Yes Yes
Blue High Yes Yes
Blue High Yes No N/A No Yes
Green Low No Yes N/A Yes Yes
Red Low Yes Yes
Comparison table of dyes for nucleus
Fig.1 Staining of nucleus in various cultured cells
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. 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.
You like to learn from
the best but missed
one of our LIVE lectures?
Sign up for
PELOAcademy and we
send you regular invitations
to our events.
Plus if you sign in for
our past webinars you
can watch the recording
are very welcome.
PELONews: Refer a friend
We are convinced that we convince you with our expertise and our outstanding service.
So if you are already a PELO-Fan, please recommend us to a colleague and
friend – you will get a coupon and nice thank-you reward. More details? Call us now.
Want to stay tuned in
and informed? Then
sign up for our regular
Reminder. Just check
the box with the infos
you like to receive and
sign up here.
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.
If you need any further
assistance or if you like
what you see, tell us:
Klopferspitz 19 82152
Planegg | Germany
Tel.: +49 89 517286
Dr. Peter Frost,
Dr Lothar Steeb