California Institute for Regenerative Medicine (CIRM) & Medical Research Council (MRC)
Human SCNT Workshop.
14 June, 2010, San Francisco, CA
Workshop report: http://www.cirm.ca.gov/files/PDFs/Publications/Human_SCNT_Workshop_Report.pdf
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Derivation of highly enriched cultures of differentiated cells from human parthenogenetic stem cells
1. Derivation of highly enriched cultures of differentiated cells from human parthenogenetic stem cells California Institute for Regenerative Medicine Medical Research Council Human SCNT Workshop 14 June, 2010, San Francisco, CA Workshop report: http://www.cirm.ca.gov/files/PDFs/Publications/Human_SCNT_Workshop_Report.pdf Nikolay Turovets, PhD Director of Research and Therapeutic Development
9. hpSC pretreatment by histone deacetylase inhibitor TSA enhances efficiency of definitive endoderm production
10. TSA pretreatment enhances efficiency of definitive endoderm production and decreases the number of remaining OCT4-positive cells
11. 2D culture system/ petri dish 3D-ECM system Other cell types in population Definitive endoderm Definitive endoderm Other cell types in population 92% 51% 49% 8% 3D-ECM differentiation system enables derivation of high-purity definitive endoderm from hpSC CXCR4 CXCR4
12. Differentiation days Developmental signaling Early hepatocyte progenitors derived from hpSC have appropriate hepatocyte morphology, and start expression of AFP and albumin hpSC Definitive endoderm (DE) derivation DE-specification toward hepatocyte fate Hepatocyte development Hepatocyte-like cells maturation 0 3 8 13 18 … Activin A Wnt3A FGF4 BMP2 HGF OSM Dex
23. ISCO Agapova Larissa Turovets Irina Agapov Vladimir Kochetkova Olga Acknowledgments Definitive endoderm ViaCyte/Novocell Ed Baedge Kevin D’Amour Mark Moorman Hepatocyte-like cells University of Pittsburg Stephen Strom RPE UC Irvine Hans Keirstead Magdalene Seiler Gabriel Nistor Funding Research was funded by ISCO and listed collaborators
Editor's Notes
The major activity of hepatocyte-like cells derived from hpSC – to restore to an OTCD patient’s missing liver function, specifically functioning urea cycle. Hepatocyte-like cells derived from hpSC could become a valuable source of cells for transplantation therapy of metabolic liver desorders caused by genetic defect: The direct differentiation procedure allow the derivation of unlimited numbers of high-purity hepatocyte-like cells from hpSC. Differentiated derivatives of hpSC may overcome limitations brought by histocompatibility issues because one hpSC line can be HLA matched with significant segments of the human population. Autologous cells cannot be used for treatment OTCD patients because they carry the same genetic defect, allogenetic HLA-matched hpSC-derived cells may overcome this “genetic defect” issue. The direct differentiation procedure allows the derivation of immature cells (progenitors) that have several advantages compared to adult liver cells: an increased proliferative capacity and plasticity; less immunogenicity; potentially superior adaptation and integration capacity; and a greater resistance to cryopreservation and ischemia.