The functional and molecular characterisation of human embryonic stem cell-derived insulin-positive cells compared with adult pancreatic beta cells
CL Basford, KJ Prentice, AB Hardy, F Sarangi, SJ Micallef, X Li, Q Guo, AG Elefanty, EG Stanley, G Keller, EM Allister, MC Nostro, MB Wheeler
DIABETOLOGIA | SPRINGER | Published : 2012
AIMS/HYPOTHESIS: Using a novel directed differentiation protocol, we recently generated up to 25% insulin-producing cells from human embryonic stem cells (hESCs) (insulin(+) cells). At this juncture, it was important to functionally and molecularly characterise these hESC-derived insulin(+) cells and identify key differences and similarities between them and primary beta cells. METHODS: We used a new reporter hESC line with green fluorescent protein (GFP) cDNA targeted to the INS locus by homologous recombination (INS (GFP/w)) and an untargeted hESC line (HES2). INS (GFP/w) allowed efficient identification and purification of GFP-producing (INS:GFP(+)) cells. Insulin(+) cells were examined f..View full abstract
Awarded by CIHR
We would like to thank A.M.J. Shapiro and T. Kin at the Clinical Islet Laboratory, University of Alberta (Edmonton AB, Canada) for the human islets used in this study. We would also like to thank R. Wang for helpful discussions and the gift of MMP2 and CK19 antibodies. We would also like to thank K. Seergobin at the Centre for Biological Timing and Cognition, University of Toronto, ON, Canada for his help and expertise in confocal microscopy, and J. Stewart at the Ontario Cancer Institute, Toronto for assistance with the transplantations. This work was supported by a grant from CIHR MOP-111241 to M.B. Wheeler. C.L. Basford was supported by a BBDC postdoctoral fellowship. K.J. Prentice was supported by a BBDC-University Health Network graduate award. M.C. Nostro was supported by a Postdoctoral Fellowship from the McEwen Centre for Regenerative Medicine. S.J. Micallef, X. Li, A.G. Elefanty and E.G. Stanley are supported by the Australian Stem Cell Centre (ASCC), The National Health and Medical Research Council (NHMRC) of Australia and The Juvenile Diabetes Research Foundation.