Journal of Tissue Engineering and Reconstructive Surgery ›› 2024, Vol. 20 ›› Issue (5): 507-.

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Generating insulin-producing cells from human adipose-derived stem cells：Comparison of three differentiation protocols

  

  • Online:2024-10-08 Published:2024-11-25

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Abstract:

Objective The induced differentiation of adipose stem cells into insulin-secreting cells is a new strategy for the
treatment of diabetes mellitus. The efficiency of the induced differentiation schemes reported so far varies greatly. This study aims to compare the differentiation efficiency of three induced differentiation schemes. Methods Fat samples from the same batch were
collected to extract fat stem cells for passage culture， and the third-generation fat stem cells were self-aggregated into balls for
induction and differentiation in different phases （one-step method， two-step method and three-step method）. The expression
levels of key genes and proteins in the products of the three schemes at the corresponding stages were detected by PCR and
immunofluorescence， and the function of insulin secreting cells was detected by glucose stimulation of insulin secretion
experiment. Results Compared with other schemes， the gene expressions of the key transcription factors INS，NKX6.1 and
PDX1 in the insulin secreting cells induced by the three-step method were significantly increased， and there were significant
differences. Key protein C-peptide and NKX6.1/PDX1 were co-expressed in insulin-secreting cells. Functionally， the insulin
secreting cells induced by the three-step method can significantly increase insulin secretion under high glucose stimulation.
Conclusion Under the same differentiation conditions， the differentiation efficiency of adipose stem cells induced into insulin
secreting cells by the three-step method is superior to that by the two-step method and the one-step method， which provides a
basis for establishing an efficient and safe induction differentiation scheme of insulin secreting cells.

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