INS(GFP/w) human embryonic stem cells facilitate isolation of in vitro derived insulin-producing cells.

ABSTRACT:

Aims/hypothesis

We aimed to generate human embryonic stem cell (hESC) reporter lines that would facilitate the characterisation of insulin-producing (INS⁺) cells derived in vitro.

Methods

Homologous recombination was used to insert sequences encoding green fluorescent protein (GFP) into the INS locus, to create reporter cell lines enabling the prospective isolation of viable INS⁺ cells.

Results

Differentiation of INS(GFP/w) hESCs using published protocols demonstrated that all GFP⁺ cells co-produced insulin, confirming the fidelity of the reporter gene. INS-GFP⁺ cells often co-produced glucagon and somatostatin, confirming conclusions from previous studies that early hESC-derived insulin-producing cells were polyhormonal. INS(GFP/w) hESCs were used to develop a 96-wel

SUBMITTER: Micallef SJ 

PROVIDER: S-EPMC3268987 | biostudies-literature | 2012 Mar

REPOSITORIES: biostudies-literature