The benefit of such an approach is the ability to generate large numbers of cells suitable for any downstream assays and applications before final differentiation into the desired cell type in vitro or in vivo. While these findings are encouraging, several challenges remain and significant efforts have been directed towards further improvement of differentiation conditions 9, 10, 11, 12, 13, the expansion of cells at different progenitor stages 14, 15 and the purification of target cell populations 16 to obtain sufficient quantities of functional pancreatic beta cells.Īn alternative strategy previously employed is the conversion of fibroblasts towards lineage-specific proliferative progenitors in vitro 17, 18, 19.
More recently, improved differentiation protocols have been described that allow the formation of functional beta-like cells from hESCs under cell culture conditions 6, 7, 8. These hESC-derived PE cells can mature into functional beta-like cells in vivo after prolonged periods following transplantation into immunodeficient mice 3, 4, 5. Stepwise differentiation protocols have been devised to guide the differentiation of human embryonic stem cells (hESCs) 1, and more recently induced pluripotent stem cells (iPSCs) 2, into definitive endoderm, primitive gut tube endoderm, posterior foregut endoderm and pancreatic endoderm (PE). Generation of functional pancreatic insulin-producing beta cells for the effective treatment of diabetes is a key area of translational research. In summary, our study has important implications for future strategies aimed at generating high numbers of functional beta cells, which may help restoring normoglycemia in patients suffering from diabetes. Transplanted cPB cells exhibit glucose-stimulated insulin secretion in vivo and protect mice from chemically induced diabetes. A screening approach identified chemical compounds that promote the differentiation and maturation of cPE cells into functional pancreatic beta-like cells (cPB cells) in vitro.
The cPF cells and their derivatives, pancreatic endodermal progenitor cells (cPE cells), can be greatly expanded. On initial culture, converted definitive endodermal progenitor cells (cDE cells) are specified into posterior foregut-like progenitor cells (cPF cells). Here we show the conversion of human fibroblasts towards an endodermal cell fate by employing non-integrative episomal reprogramming factors in combination with specific growth factors and chemical compounds.
Endotoxin levels are verified, using the LAL method to ensure consistency for use across multiple applications. This human recombinant TGF-ꞵ1 is precisely reconstituted to 0.1 mg/ml in 100 mM acetic acid and requires no additional preparation, improving reproducibility.
Tissue and Cell Culture Dissociation ReagentsĬonveniently and consistently add transforming growth factor beta 1 (TGF-ꞵ1) to your cultures with this cell culture-ready formulation.Work at STEMCELL View Current Opportunities >
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