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Neuromics Presentation V4

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In Search of Better Tools and Capabilities
"We need to do more with less." This is a common theme we hear in our conversations and follow up with Researchers in early phase drug discovery.

To us this translates into better identification and more intense screening of potential therapeutic compounds and targets. Failure is success if it is determined before animal testing.

The foundation being potent, pure and easy to culture primary cells. Then building on the platform with the required media/growth factors, markers, transfection reagents and apoptosis detection kits.

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Neuromics Presentation V4

  1. 1. Tools and Capabilities Focus on cell based assays and gene expression analysisfor early phase Drug Discovery 3/5/2011-Version 4
  2. 2. Focus: Provide researchers a subset of tools and methods to enable successful outcomes in early phase drug discovery. Failure is success when determined early.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 Images: STEMEZ™ hNP1™ can be differentiated into various neuronal subtypes under defined conditions. DAPI GFAP Antibody • > 80% Tuj1+ • > 60% MAP2+ 3/5/2011
  3. 3. 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 3/5/2011
  4. 4. Stable, Potent and Cost Effective Cells Growth Derived and manufacturedFactors/Markers neural cells from Applications include: human embryonic stem Media/3-D cell line H9 •Proliferation and Gels/ECMS (WA09). neurogenesis •Neuronal differentiation •Neurite outgrowthGene 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.. 3/5/2011
  5. 5. Assay kits, Qualified Media and Reagents-Sample Images Markers/Antibodies Growth Factors/Culture Media Gene Expression Analysis Apoptosis DetectionImges: GFAP (Catalog#: STEMEZTMhNP1 i-Fect knockdown of Delta Opioid Jurkat cells duallyaCH22102) Staining of of stained with HoechstAstrocytes in the Ventral Human Neural Receptor. Molecular Pain 2005,horn. Courtesy of Dr. Qi-lin Progenitors treated 1:29 and PolycaspaseL. Cao, University of with hGDNF Assay Kit,Louisville School of cultured inMedicine. Rat spinal cord STEMEZ hNP1homogenate WBs: NF-H,NF-M,, NF-L,, NF66 and Neural ProgenitorGFAP. Expansion Media 3/5/2011
  6. 6. 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 3/5/2011
  7. 7. Tools and Capabilities Cell Based Assays-ExamplesSTEMEZ™ hNP1™ Human Neural Progenitor Kit is an efficient tool to quantitativelydetect both stimulators and inhibitors of cell migration when used with the OrisTM CellMigration Assay Kit (Platypus Technologies). Method development to date indicates thatthe assay has the potential for adaptation as a homogenous HTS-suitable cell-basedassay. 3/5/2011
  8. 8. 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 touse 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 wellas YFP. On the following day, these cells were infected with HeV or VSV pseudotyped virusesbearing 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 pseudotypedvirions; (C) upon excitation for YFP, and the green fluorescence shows the efficiency oftransfection in neurons. This experiment indicates that the proposed experiments can be carriedout in primary neurons, which are transfectable and infectable in our systems, and thus supportsall the proposed aims. Data Courtesy of Dr. Matteo Porotto, Weill Cornell Medical College. LargerImage 3/5/2011
  9. 9. Cell Based Assays-Examples Primary Rat Neurons and Astrocytes Data Courtesy of Evanna Gleason, Louisiana State University 3/5/2011
  10. 10.  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 3/5/2011
  11. 11. 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 3/5/2011
  12. 12.  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 3/5/2011
  13. 13.  Pete Shuster CEO and Owner Direct Phone: 612-801-1007 www.neuromics.com pshuster@neuromics.com www.neuromics.net neuromics.blogspot.com Social Networking: sirnatransfection.blogspot.com 3/5/2011

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