Biopharma musculoskeletal disorders_draft 6-30-2013 jm edits
Harnessing the Power ofCellsTMNew Assays forMusculoskeletal Disorders DrugDiscoveryExecutive Overview1
Solutions for Musculoskeletal Disease ResearchPotent Umbilical Cord Blood Derived Human Mesenchymal Stem Cells Alternative to induced pluripotent and embryonic stem cells Phenotypically and morphologically stable through 50 population doublings HLA-DR-, CD14-, CD19-, CD31-, CD34-, CD41a-, CD235a-, ALP-, CD271-CD45-, D7FIB++++, CD44++++, CD54++++, CD73++++, CD90++++,CD105++++, CD140b++++, CD166++++, CD146++++ Capacity to produce cell numbers required for high throughput & contentscreeningDifferentiated Cells: Osteoblasts,Chondrocytes and FibroblastsPotent Media Low serum and clinical grade, serumfree expansion media Osteogenesis, chondrogenesis &adipogenesis media Stem cell basal mediaMSCGroTM Media vs Lonza1 6-1-2013 2
Potent Cells and Media-Better Results=Lower Costs2 4/30/2013 3Our Media Keeps your Cells Going and Going “We tested theeffects of MSCTMGro defined medium using several differentlots of human adult primary stem cells and found thatMSCGro supports a more robust proliferation rate thannormal undefined media. This provided shorter doublingtimes and increased cellular yield, and maintained the cells inan undifferentiated state. We also found that MSCGromedium is stable under normal laboratory conditions for anextended time period compared to other defined media.” BenBuehrer, VP and CSO, Zen-BioImages: (A) Human cord-blood MSCs were expanded inlow-serum MSC-GroTM to confluence as shown here. (B)were differentiated in osteogenic MSC-GroTM. Early stageosteoblasts are shown here; the arrow shows earlyformation of mineralized matrix. (C)&(D) Matureosteoblasts stained positive for Alizarin red. Phase contrastimage at 200 x, scale bar is 50 mmeters.
Potent Cells and Media-Better Results=Lower Costs2 4/30/2013 4Human FibroblastsImages: Pancreatic stellate cells are positive for GFAP and a -SMA.Immunofluorescence analysis of representative primary PSC lines grown onchamber slides. Cells were stained with 40, 6-diamidino-2-phenylindole (DAPI;blue) and antibodies (green) for vimentin, GFAP, and alpha-SMA. The humanPANC-1 cell and HPF line were negative controls for alpha-SMA staining. Slideswere visualized at 40 magnification. Published OnlineFirst March 20, 2013; doi:10.1158/0008-5472.CAN-12-4601. Cancer Res May 15, 2013 73; 3007.Figure: Continuous expansion of human fibroblasts.Cells expanded through 16 passages. Fibroblastwere plated at 5,000 cells/cm2 in Greiner Bio-oneT25 flasks maintained in VitroPlus III, low serummedia in a reduced oxygen environment (5% O2, 5%CO2, 90% N2) at 37OF in a humidified chamber. Thepancreatic fibroblasts were detached using byincubation in AccutaseTM they were approximately at80-90% confluency and counted using a Beckman-Coulter Z2.
Potent Cells and Media-Summary3 4/30/2013 5Cells are differentiated from human MSCs-Competitive to IPSCs since there is noreprogramming nor epigenetic issues intrinsic toIPSCsCost-effective due to manufacturing scalabilityDirect purchase without licensing requirementHighly experienced team with extensive technical andbiopharma partnership experience.Cell therapy application as well as discovery.
Capabilities meet HCS/HTS Requirements Cost-effective materials to meet target per well costs Human cells reduce false positives vs. animal tissues Cells express proteins of interest. Verification assays providedbased on specific requirements. Readily scalable: Identical phenotype/genotype from pilot toscreening mode. Co-culture capability for custom assays. E.g., chondrocyte/osteoblasts4 4/30/2013 6
Evaluation Process Confidential discussion of Specific Disorder, Therapeutic Target(s) and cellbased assay requirements. Design pilot assay with deliverables and milestones Review milestone data vs requirements Revise assay as needed Determine final assay configuration. Determine scale up requirements Cost targets Material and deliverables Evaluate progress vs plans and adjust accordingly Position capabilities for future projects5 4/30/2013 7
ContactsTechnical and Consulting:Dr. Jim MusickPresidentVitro Biopharmajim@vitrobiopharma.com(303) 999-21304621 Technology DriveGolden, CO 80403Sales/Proposal ProcessPete ShusterCEONeuromicspshuster@neuromics.com612-801-10075325 West 74ths StreetEdina, MN 55439Draft 1 04-2013-3 8