PeloNews Feb/March2019

PELONEWS
WITH FOCUS ON 3D INNVOATIONS
PELOBiotech Newsletter March 2019
“Vita brevis, ars longa.“
(Seize the day.)
Dear Scientist,
Dear Scientist,
The UK is due to leave the European Union on Friday 29th March 2019,
after the UK voted to leave in the 2016 referendum.
Understandably you will be considering the changes it may have on
your supply deliveries. PELOBiotech GmbH as your trusted distributor
has prepared for all potential Brexit outcomes and is committed to
supporting you during Brexit and to manage the supply chain
risks in these challenging conditions.
We have made extensive preparations to ensure continuity of service
during and post-Brexit. Our core capability is clarifying and optimising
global supply chains across international borders and managing regulatory
complexity. Any new regulatory requirements will be integrated into
our “ready to send” process so we can minimizedthe risk of customs
delays at the border due to paperwork discrepancies. We will continue
to keep you updated as appropriate. In the meantime, to discuss how
we can support your Brexit preparations, please contact us anytime.
Cheers. Best regards,
Christiane Büchsel
Editor in Chief PELONews
Hippokrates, Founder of Medicine
as Science-
Content
PELOAcademy: Learn
from the best
Thank you alll
Meet us at…
Paper Altert
Featured Fields
&Products
3D
BBB
Cancer
Chimerism Assay
Microfluidic Chambers
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From Bedside to Bench: Translating
Clinical Tumour Treatments into 3D Culture
'Translational research' is mainly seen as the effort to develop basic research
into clinical routine, this webinar shows how 3D culture systems
may serve as a comprehensive tool to reproduce patient treatment protocols
in your laboratory.
From a lab perspective, the clinical situation looks amazingly simple:
Both patients and doctors do not bother much about biological endpoints
as apoptosis or DNA damage of cancer cells, but focus on general ones:
Does the tumour shrink? And finally: Is the tumour cured?
Applying such 'simple' endpoints on 3D tumour cell cultures can make a
lot of sense:
Volume response as a readout summarises all anti-tumour effects
into one
Sign up here
Watch now here
In long-term cultures, each single micro-tumour may be assigned to
be either 'cured' or 'resistant'
The concept of clonal evolution requires time and large 'cohorts" of
micro-tumours. Size-based screening is an efficient method to detect
specific mutations and other rare events. In this webinar, you will
learn:
How to design and apply complex treatment schedules to microtumours,
as chemoradiation, fractionated radiotherapy and hyperthermia
treatment •
How to handle cancer cell types that normally do not grow as spheroids
How optical scanning and paraffin sections are easily used to assess
treatment response
How to detect and isolate micro-tumours that did not respond to treatment
Watch now this great vivid lecture by Dr. Thomsen.

Advanced in-vitro Models for Tumor Models
presented by Tommaso Sbrana, CEO IVtech, Italy
A standard 2D static in-vitro models is not representative of the reality.
This is well known and it is more than accepted if the goal is the estimation of an in-vitro model to mimic
a tumor environment. The lack of crosstalk between tissues as well as the static cell culture conditions
are limitations in the correlation between the standard 2D in-vitro model and the scenario that we desire
to reproduce. It is required an update of the technology used to set up an in-vitro model, in order to be
able to recreat the pato-physiological stimuli, increasing its predictivity of the human scanario. Therefore
IVTech offers an enabling technology, represented by advanced cell culture chambers, to permit the study
of a 3D dynamic in-vitro model, characterized by a crosstalk between different tissues.
Using these products, it is now possible to add the IVth dimension (the dynamicity) and the
Vth dimension (the crosstalk between different tissues) to a 3D in-vitro model. The focus is shifted from
a single tissue to a pathway evaluated in dynamic conditions. The use of this chambers in oncology for
instance allows to develop advanced in-vitro models to mimic the tumor environment, test the effects of
an antitumor compound as well as evaluate the interactions between healthy and disease tissues.
Therefore it is possible to:
recreat the tumor micro-environment and its main characteristics
(i.e. hipoxia )
tissues, in order to study the uptake mechanism
investigate the tumor cell resistance to a drug exposure, as it is
sometimes observed in human (i.e. brest cancer applications)
Watch now here
evaluate the interactions between floating metastasis and healthy
study the new frontiers of tumor treatments represented by immunotherapy, combining tumor and limphatic
models. Watch now this fantatstic webinar by Tommaso.Sbrana.

PELONews
Let‘s talk
Optimised
Cancer
Treatments
3D Models
Symposium Munich:
Wed, 27 March 2019
Meet high end experts in the field
of 3D cancer models and treatments.
Learn what they are doing
and how they have optimized
their treatments.
3D CoSeedis: a novel and innovative
3D microwell array in the
analysis of adhesion independent
micro-organoids
3D CoSeedis consists of a unique
conical agarose matrix array that
allows the formation of spheroidal
and non-spheroidal cell aggregates
in a highly reproducible and consistent
manner. Furthermore, 3D
CoSeedis allows the supporting
growth of feeder cells in the formation
of 3D cell constructs that are
not in direct contact with the test
cells themselves. Validated protocols
and workflows have been developed
to analyse fully formed 3D
aggregates.
In summary, the modular and standardised
3D CoSeedis systems
broaden the number of cell types
accessible to examination in 3D
substantially and present a versatile,
robust and reliable platform for
3D cell culture studies.
Sign up now at
info@pelobiotech.com and let
Katja and Christiane know that you
will be part of this groundbreaking
Symposium.

New Products
SynVivo 3D Tissue/Organ on Chip Models
SynVivo is a cell-based microfluidic platform that provides a morphologically and biologically realistic microenvironment
allowing real-time study of cellular behavior, drug delivery and drug discovery. Watch video showing
SynVivo in action.
SynVivo has developed 3D tissue models that recreate complex in vivo microvasculature including scale, morphology,
hemodynamic shear stress and cellular interactions with side-by-side architecture enabling real-time
visualization. 3D tissue models include SynBBB (blood brain barrier) SynTumor
(Cancer), SynRAM (Inflammation) and SynTox (Toxicology). The platform can be adapted to replicate the
unique features of any desired organ and tissue for your specific applications.
SynBBB in vitro blood brain barrier (BBB) model allows visualization of cellular interactions and barrier functionality
in real time. Evaluate and quantitate permeability of therapeutics and small molecules across the endothelium
of the BBB in real-time. Unlike BBB models that are membrane-based and arranged in top to bottom
architecture (i.e., Transwell), small molecule transport can be visualized, accessed and quantified in real-time
using the SynBBB model due to its side-by-side architecture.
Human or rat species in co-culture with endothelial cells and astrocytes or tri-culture to include pericytes can
be used to assess the passage of molecules through the BBB and the effect of molecules on the BBB physiology
(normal or diseased). Various types of molecules can be assessed such as nanoparticles, small
molecules, peptides or large molecules including antibodies. Cellular interactions including vascular - immune
cell interactions can be studied in this model.
Apical chamber (outer blue channels) is for culture of
vascular cells while the basolateral chamber (central
red chamber) is for culture of brain tissue cells
(astrocytes, pericytes) which can be mixed or separated
in different compartments. Engineered porous architecture
enables communication between the vascular
and tissue cells.
F
The SynBBB model is available as kits or microfluidic chips.
Publications using the SynBBB model
(1) A Microfluidic model of human brain (uHuB) for assessment of blood brain barrier.
Tyler D. Brown, Maksymillian Nowak, Alexandra V. Bayles, Balabhaskar Prabhakarpandian, Pankaj Karande, Joerg Lahann,
Matthew E. Helgeson and Samir Mitragotri. BioEngineering and Translational Medicine 2019; 1-13.
(2) Protein kinase C-delta inhibition protects blood brain barrier from sepsis-induced vascular damage.Yuan Tang, Fariborz
Sorush, Shuang Sun, Elisbetta Liverani, Jordan Langston, Qingliang Yang, Laurie Kilpatrick and Mohammad Kiani. Journal of
Neuroinflammation 15:309 (2018)
(3) Trastuzumab Distribution in an In-Vivo and In-Vitro Model of Brain Metastases of Breast Cancer Tori B. Terrell-Hall, Mohamed
Ismail Nounou, Fatema El-Amrawy, Jessica I.G. Griffith and Paul R. Lockman Oncotarget. 2017; 8:83734-83744
Learn more
(4) Permeability across a novel microfluidic blood‐tumor barrier model. Tori B. Terrell‐Hall, Amanda G. Ammer, Jessica I.
G. Griffith and Paul R. Lockman Fluids and Barriers of the CNS (2017) 14:3
(5) A Novel Dynamic Neonatal Blood-Brain Barrier on a Chip. S. Deosarkar, B. Prabhakarpandian, B. Wang, J.B. Sheffield,
B. Krynska, M. Kiani. PLOS ONE, 2015

PELONews
SynBBB exhibits significant tight junction formation and shows excellent correlation
with in vivo permeability
Other validated tissue models

Chimerism analysis: Non-T & Non-B Genomic Detection Kits
· Q: Is my current sorting method compatible with these kits?
A: Yes! The Non-T Genomic and Non-B Genomic Detection Kits determine cell purity at the nucleic
acid level. It is therefore insensitive to the method used to sort the cells.
Q: Do I need to introduce a new technique and procedures in the lab to use these kits?
A:These kits are designed to be used along the traditional chimerism analysis workflow. All there is to
do is add the DNA sample to the reaction tube. Then the reaction can be run at the same time than the
chimerism PCR, because the reaction parameters are the same. The PCR products are analyzed on a
sequencer in the same way than the chimerism sample. Chimerism labs already have all the expertise
and equipment to run the Non-T Genomic and Non-B Genomic Detection Kits.
Q: I don't have a lot of DNA sample to start with, I need it all for the chimerism analysis and
cannot waste any. Will I have enough sample to check for purity?
A:The Non-T Genomic and Non-B Genomic Detection Kits can be run with as little as 0.5 ng of genomic
DNA. This corresponds to the amount of DNA present in approximately 200 cells. The lowest DNA
concentration that can be detected with classical UV methods is about 2 ng/µL. So even with samples
with very low, non detectable DNA content, 1 µL of sample is usually sufficient to run the Genomic Detection
Kit.
Learn more here
Q: I validated my cell sorting method and had good purities. Why would I need to check the
purity of the sorted cells every time?
A:The quality of the cell sorting is very sample-dependant. The first source of viability is of course the
patient. Shortly after hematopoietic stem cell transplant, it is common for patients to have a very low
blood cell count, as well as a very low T cell count (often less than 5% of total leukocytes, whereas
normal levels are 25-30%). The risk of T Cells isolated from these samples to be contaminated by monocytes
or granulocytes is very high. In addition, blood samples sometimes reach the lab several days
after collection, and can be exposed to extreme temperatures during transportation. These parameters
can vary between samples, are out of the lab's control, and can have a very negative impact on cell
sorting quality. Assessing the purity of each isolated cell subset is an essential
quality control step, and is required for ASHI or EFI accreditation.
Learn more
Q: Isn't flow cytometry a more accurate method of assessing purity?
A: Actually, flow cytometry is not as accurate as one might think, because measured purity can vary
considerably depending on the way the data is analyzed. On most samples, changing the way the cells are
gated can lead to purities readings anywhere between 80% and 100%. In contrast, calculating purity is
straightforward with the Non-T Genomic and Non-B Genomic Detection Kits, and there is no interpretation
or ambiguities during the analysis.
In addition, chimerism tests are based on DNA analysis, and so are the Non-T Genomic and Non-B Genomic
Detection Kits. All the DNA present in the sample will be taken into account. In contrast, flow cytometry
analysis excludes "debris" and events that have a small size. These may contain DNA fragments or even
entire nuclei. This DNA, excluded from the flow cytometry data, might have an impact on the chimerism
analysis results.

PELONews
PELONews: Refer a friend
We are convinced that we convince you with our
expertise and excellent service. So if you are
already a PELO-Fan, please recommend us to a
colleague and friend – you will get a nice thankyou
personalized reward. More details? Call us
now.
Thank you all: Here‘s a quote from our Customer-survey: 100% are very satisfied
or satiesfied with PELOBiotech!! You are wonderful and we are happy to
serve you..
Paper Alert Our partner ReSyn has publisehd a new paper.
„Immune genes are primed for robust transcription by proximal long noncoding
RNAs located in nuclear compartments.“
by Stoyan Stoychev and his scientific partners. Nature Geneticsvolume
51, pages138–150 (2019) |
Accumulation of trimethylation of histone H3 at lysine 4 (H3K4me3) on immune
-related gene promoters underlies robust transcription during trained immunity.
However, the molecular basis for this remains unknown. Here we show threedimensional
chromatin topology enables immune genes to engage in chromosomal
contacts with a subset of long noncoding RNAs (lncRNAs) we have defined
as immune gene–priming lncRNAs (IPLs). We show that the prototypical
IPL, UMLILO, acts in cis to direct the WD repeat-containing protein 5 (WDR5)–
mixed lineage leukemia protein 1 (MLL1) complex across the chemokine promoters,
facilitating their H3K4me3 epigenetic priming. This mechanism is
shared amongst several trained immune genes. Training mediated by β-glucan
epigenetically reprograms immune genes by upregulating IPLs in manner dependent
on nuclear factor of activated T cells. The murine chemokine topologically
associating domain lacks an IPL, and the Cxclgenes are not trained. Strikingly,
the insertion of UMLILO into the chemokine topologically associating
domain in mouse macrophages resulted in training of Cxcl genes. This provides
strong evidence that lncRNA-mediated regulation is central to the establishment
of trained immunity.
Want to meet Stoyan and talk about
what ReSyn can do for you:
EUPA 24-28th March, Potsdam.
Meet also our sales expert Katja
Keystone Symposium, 7-11th
April, Stockholm, Sweden.
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.
Save these
dates
Let`s talk:
Optimised Cancer
Treatments with 3D
Models
March 27, 2019
10 AM ct – 1PM
Sky Lounge G2B
Club im IZB
IGLD: 07.- 09.
March 2019, Frankfurt
Elrig: 07. March
2019, Darmstadt
ADF: 13.- 16.March
2019, Munich
EUPA 24-28th
March, 2019, Potsdam
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