Tools and Capabilities

Focus on cell based assays

and gene expression analysis
for early phase Drug Discovery

12/08/2021-Version 4
Focus:
 Provide researchers a subset of tools and methods to enable successful
outcomes in early phase drug discovery. Failure is success when
determined early.*
*90% of lead candidates identified by current in vitro systems fail to become drugs

Value (simplified):
 Physiologically relevant primary human (STEMEZ™), rat and mouse
cells
 Better in vitro screening of targets and molecules =
 Reduced animal model use
 Shorten development timeline
 Lower downstream costs
TUJ DAPI
TUJ
Images: STEMEZ™ hNP1™ can be
differentiated into various neuronal
subtypes under defined conditions.
DAPI
GFAP Antibody
TUJ DAPI DAPI DAPI • > 80% Tuj1+
Hb9 ChAT TUJ GABA
DAT TUJ • > 60% MAP2+

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Why Neuromics?
Standing out in a crowded room

 We have a experience serving many large Pharmas, Biotechs,

Academic and Government Labs.

 Customer success focused

 Expert technical support
 Detailed protocols and methods
 Quick replacements and reorders
 Large expert resource network
 Published expert input
 Customer feedback and data sharing through
social media outlets and blogs

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Tools and Capabilities
Stable, Potent and Cost Effective Cells

Growth Derived and

manufactured
Factors/Markers neural cells from Applications include:
human
embryonic stem
Media/3-D cell line H9 •Proliferation and
Gels/ECMS (WA09). neurogenesis
•Neuronal differentiation
•Neurite outgrowth
Gene Expression •Cell migration
Analysis •Neurotoxicity screening
•Cell based compound
Apoptosis screening
Detection •G-protein coupled receptors
•Ion channels
Cell Migration •Stem cell transplantation
Assays

Testimonial: got 10 million cells total after extraction from the tissue.  At Day 4 they all  developed long axons.
Thank you so much for the replacement."  Dr. Lidia Gardner, University of Tennessee HSC..

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 Tools and Capabilities
Assay kits, Qualified Media and Reagents-Sample Images
Markers/Antibodies
Imges:  GFAP (Catalog#: a
CH22102) Staining of of
Astrocytes in the Ventral
horn. Courtesy of Dr. Qi-lin
L. Cao, University of
Louisville School of
Medicine. Rat spinal cord
homogenate WBs: NF-H,
NF-M,, NF-L,, NF66 and
GFAP.
Growth Factors/Culture Media Gene Expression Analysis Apoptosis Detection
STEMEZTM i-Fect knockdown of Delta Opioid Jurkat cells dually
hNP1 Human Neural
Receptor. stained with Hoechst
Progenitors treated with
Molecular Pain 2005, 1:29  and Polycaspase
hGDNF cultured in
STEMEZ hNP1 Neural Assay Kit,
Progenitor Expansion
Media
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Tools and Capabilities
Cell Based Assays-Examples
 STEMEZ hN2 Human Neural Discovery Kit

 Phenotyping
Neural phenotypes derived from N2 cell
lines. (A) Phase contrast image of
differentiated culture. (B) Network
including post-mitotic motoneurons
(HB9). (C) Cholinergic neuron. (D) Tuj-1
positive cells that are DAT-positive
(dopamine transporter; closed arrow)
and DAT-negative (open arrow). (E)
Gabaergic neurons, inset illustrates
GABA in axon, but not the dendrites
(arrow).

 Electrophysical and FLIPR Data Provided upon request

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Tools and Capabilities
Cell Based Assays-Examples

STEMEZ™ hNP1™ Human Neural Progenitor Kit is an efficient tool to quantitatively

detect both stimulators and inhibitors of cell migration when used with the Oris TM Cell
Migration Assay Kit (Platypus Technologies). Method development to date indicates that
the assay has the potential for adaptation as a homogenous HTS-suitable cell-based
assay.

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Tools and Capabilities
Cell Based Assays-Examples
 Primary Rat Neurons and Astrocytes
Transfection of functional HeV
glycoproteins and infection with HeV
pseudotyped virions.In order to establish
the feasibility of carrying out the proposed
experiments in primary neurons, we show
(figure ) that our assays are amenable to
use in primary neurons.  In the experiment, Combined Hippocampus, Cortex, and Ventricular -E18
(Neuromics) were plated, and at 3 days were transfected with plasmids encoding HeV G/F as well
as YFP.  On the following day, these cells were infected with HeV or VSV pseudotyped viruses
bearing RFP.  In the figure, (A) the phase contrast photos show the differentiated neurons; (B)
upon excitation for RFP, the red fluorescence indicates neurons infected by HeV pseudotyped
virions; (C) upon excitation for YFP, and the green fluorescence shows the efficiency of
transfection in neurons.  This experiment indicates that the proposed experiments can be carried
out in primary neurons, which are transfectable and infectable in our systems, and thus supports
all the proposed aims. Data Courtesy of Dr. Matteo Porotto, Weill Cornell Medical College.
Larger Image 

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Tools and Capabilities
Cell Based Assays-Examples
 Primary Rat Neurons and Astrocytes

Data Courtesy of Evanna Gleason, Louisiana State University

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Tools and Capabilities

 Gene Expression Analysis

 It’s all about delivery!
 i-FectTM siRNA delivery kit : Success in delivering siRNA to
DOR, hTERT, The β3 subunit of the Na+,K+-ATPase,
rSNSR1, NTS1. NAV1.8, Survivin,  TRL4, Flaviviruses and
more.
 Tools for delivering plasmids, proteins, antisense and
other small RNA and DNA molecule.

 Transfection Delivery: Customer Publications

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Tools and Capabilities

More Key Product Category Links:

 Antibodies-Markers

 Apoptosis Research Reagents -includes:

detection kits, antibodies and proteins

 Proteins Recombinant and natural proteins-

includes a wide selection of cell growth factors.
 Transfection Reagents-Gene Expression Analysis

 Cell Growth Media

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Next Steps
 Clear Understanding of where Neuromics could have
a positive impact.
 More intense in vitro screening of compounds and
targets.
 Better candidates for downstream processes.

 Propose solutions based on this understanding

 Follow up, tweak and improve

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 Pete Shuster Neuromics Website and
CEO and Owner Blogs:
Direct Phone: 612-801-1007 www.neuromics.com
News Behind the Neuroscience
pshuster@neuromics.com News blog: www.neuromics.net
Methods and pubs blog:
neuromics.blogspot.com
 Social Networking:
Transfection blog:
sirnatransfection.blogspot.com
  

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