Differentiation of insulin-producing cells from human umbilical cord mesenchymal stem cells infected by MAFA-PDX1 overexpressed lentivirus

doi:10.12307/2024.104

Abstract

Abstract: BACKGROUND: Transplantation of stem cell-derived islet β cells has been considered effective for the treatment of type 1 diabetes. Human umbilical cord mesenchymal stem cell is an ideal cellular source, but with a low differentiation efficiency to islet β cells.
OBJECTIVE: To explore the possibility of human umbilical cord mesenchymal stem cells modified by MAFA and PDX1 to differentiate into insulin-producing cells.
METHODS: MAFA-PDX1 lentivirus expression vectors were constructed. The efficiency and potentiality of human umbilical cord mesenchymal stem cells differentiated into insulin-producing cells with three methods were compared by cell morphology, RT-qPCR, and dithizone staining [protocol A: Simple lentivirus group; protocol B: Drug (nicotinamide β-mercaptoethanol) induction followed by lentivirus group; protocol C: lentivirus and drug induction group].
RESULTS AND CONCLUSION: (1) Morphological change of cells: Cell morphology was all altered after the induction of three protocols. At day 11, human umbilical cord mesenchymal stem cells induced by protocol B showed the most cell clusters among the three protocols, appearing aggregated islet-like cell clusters. (2) Islet-related gene expression detected by RT-qPCR: Horizontal comparison of the three protocols at the same induction time point showed that the expression levels of MAFA and PDX1 genes were the highest in protocol C on day 5 of induction, and those in protocol B were the highest on day 11 of induction. Human umbilical cord mesenchymal stem cells induced by protocol B had the greatest expression of GCG gene at day 5, INS and GLUT2 genes at day 11. (3) Dithizone staining to identify zinc ions: parts of the post-induced cells were stained brownish red by dithizone on day 11. The partial small island cells were stained brownish red with a darker color (positive expression) in protocol B. (4) It is concluded that the overexpression of MAFA and PDX1 can promote the differentiation of human umbilical cord mesenchymal stem cells into insulin-producing cells. The combination of MAFA-PDX1 gene modification and drug induction is superior to the single gene modification.

Key words: umbilical cord mesenchymal stem cell, MAFA, PDX1, lentivirus overexpression vector, insulin-producing cell, diabetes, induced differentiation

CLC Number:

Qiu Xiaoyan, Li Bixin, Li Jingdi, Fan Chuiqin, Ma Lian, Wang Hongwu. Differentiation of insulin-producing cells from human umbilical cord mesenchymal stem cells infected by MAFA-PDX1 overexpressed lentivirus[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(7): 1000-1006.

Figures/Tables 11

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