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HNF-6-independent differentiation of mouse embryonic stem cells into insulin-producing cells. Paper-10015825. METHODS: We generated mouse Hnf6-/- embryonic stem cells and grew them as embryoid bodies.
RESULTS: No difference was observed in the expression of insulin between wild-type and Hnf6-/-embryoid bodies.
In both cases insulin was expressed in the outer layer of cells, which is similar to the visceral endoderm.
The expression of genes that are targets of HNF-6 in developing pancreas was unaffected in Hnf6-/-embryoid bodies. Paper-10015825.
In wild-type embryoid bodies HNF-6 was transiently expressed in the outer layer of cells, but was not co-expressed with insulin. Paper-10015825.
To achieve such optimization, one needs to know whether the differentiation of cells in embryoid bodies mimicks that of pancreatic beta cells in embryos.
The expression of HNF-6, insulin, and transcription factors that are regulated by HNF-6 in developing pancreas was compared in wild-type and Hnf6-/- embryoid bodies. Paper-10015825.
CONCLUSION/INTERPRETATION: In contrast to the development of pancreatic beta cells, the differentiation of insulin-producing cells in embryoid bodies did not require HNF-6. Paper-10015825.
Thus, the differentiation mechanism of insulin-producing cells in embryoid bodies differs from that of the beta cells and it is likely to resemble that of insulin-producing cells in the visceral endoderm.
AIMS/HYPOTHESIS: Embryonic stem cells, when grown as embryoid bodies, spontaneously generate insulin-producing cells which could be used in therapy of diabetes mellitus, provided that their selection and differentiation are optimized.
To address this question we verified if the differentiation of the insulin-producing cells in embryoid bodies requires Hepatocyte Nuclear Factor-6 ( HNF-6), a transcription factor known to control pancreatic endocrine differentiation in embryos. Paper-10015825.
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