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Author information

Dr. H.J. Paek
Center for Biomedical Engineering, Brown University, Providence, Rhode Island.
Department of Molecular Pharmacology, Physiology and Biotechnology, Brown University, Providence, Rhode
L.J. Moise
Department of Molecular Pharmacology, Physiology and Biotechnology, Brown University, Providence, Rhode
J.R. Morgan
Center for Biomedical Engineering, Brown University, Providence, Rhode Island.
Department of Molecular Pharmacology, Physiology and Biotechnology, Brown University, Providence, Rhode
M.J. Lysaght
Center for Biomedical Engineering, Brown University, Providence, Rhode Island.
Department of Molecular Pharmacology, Physiology and Biotechnology, Brown University, Providence, Rhode

ABSTRACT

Islet-like cell clusters (ILCCs) were derived from murine embryonic stem cells using a slightly modified version of the protocol originally described by Lumelsky et al. in 2001. Analysis with enzyme-linked immunosorbent assays (ELISAs) that distinguish human from murine insulin demonstrated that insulin released from these ILCCs, upon initial in vitro glucose challenge, was of non-murine origin and in fact corresponded to the species of insulin, human or bovine, that had been added to the culture media used to derive ILCCs. This finding convincingly supports the hypothesis that ILCCs are not synthesizing insulin de novo, but rather simply regurgitating insulin taken up during tissue culture. In further experiments, ILCCs were derived in media in which insulin had been replaced by IGF-I with which it shares a common signaling pathway. These ILCCs failed to release any detectable insulin. In contrast, ILCCs produced by various protocols stained positive (dithizone and immunoselective antibodies) for intracellular insulin and, in some cases, C-peptide. Despite the presence of at least some level of de novo, synthesized insulin in ILCCs, the majority of insulin released by ILCCs was sequestered from the exogenous medium.

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PDF Document ImageOrigin of Insulin Secreted from Islet-Like Cell Clusters Derived from Murine Embryonic Stem Cells

Dr. H.J. Paek, L.J. Moise, J.R. Morgan, and M.J. Lysaght

Cloning and Stem Cells. December 2005, Vol. 7, No. 4: 226-231

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