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Title: Generating mESC-derived insulin-producing cell lines through an intermediate lineage-restricted progenitor line
Authors: Li, G. 
Luo, R.
Zhang, J. 
Lian, Q. 
Xie, F.
Lim, S.K. 
Salto-Tellez, M.
Yeo, K.S.
Tan, E.K.W.
Caille, D.
Meda, P.
Kon, O.L.
Issue Date: 2009
Citation: Li, G., Luo, R., Zhang, J., Lian, Q., Xie, F., Lim, S.K., Salto-Tellez, M., Yeo, K.S., Tan, E.K.W., Caille, D., Meda, P., Kon, O.L. (2009). Generating mESC-derived insulin-producing cell lines through an intermediate lineage-restricted progenitor line. Stem Cell Research 2 (1) : 41-55. ScholarBank@NUS Repository.
Abstract: Generating surrogate insulin-producing cells from embryonic stem cells (ESCs) through in vitro replication of successive steps during pancreatic development has been challenging . Here we describe a novel reproducible protocol to establish homogeneous and scalable insulin-producing cell lines from mouse (m) ESCs via differentiation of the previously described lineage-restricted clonal mESC-derived E-RoSH cells. Unlike their parental mESCs, E-RoSH cells expressed high levels of mesodermal and endodermal genes. Nutrient depletion in the presence of nicotinamide inhibited proliferation of E-RoSH cells and induced differentiation into heterogeneous cultures comprising vascular-like structures that produced detectable levels of insulin and C-peptide in an equimolar ratio. Limiting dilution of these cultures resulted in the isolation of eight independent insulin-producing cell lines in five experiments. All these lines were cloned and shown to be amenable to repeated cycles of freeze and thaw and to replicate for months with a doubling time of 3-4 days. Under such conditions, the cultured cells exhibited genomic, structural, biochemical, and pharmacological properties of pancreatic β cells, including storage of an equimolar ratio of insulin and C-peptide in granules and release of the contents of these organelles through a glucose-sensitive machinery. After transplantation, these cells reversed hyperglycemia in streptozotocin-treated SCID mice and did not form teratomas. © 2008.
Source Title: Stem Cell Research
ISSN: 18735061
DOI: 10.1016/j.scr.2008.07.006
Appears in Collections:Staff Publications

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