Department of Medicine and Therapeutics, The Chinese University of Hong Kong, The Prince of Wales Hospital, Shatin, Hong Kong SAR, China.
Cell Transplant. 2013;22(1):147-58. doi: 10.3727/096368912X653057. Epub 2012 Jul 5.
The use of pancreatic β-cells differentiated from embryonic stem (ES) cells or induced pluripotent stem (iPS) cells is a promising strategy in cell therapy. Pancreatic β-cell development is regulated by the sequential expression of a molecular network of transcription factors. In this experiment, we adopted a three-step differentiation protocol to differentiate mES (mouse ES) cells into insulin-secreting cells and overexpressed transcription factors by adenoviral vectors at various combinations at different time of differentiation. We found that the coexpression of Pdx1 and MafA with either Ngn3 or NeuroD, especially at the final stage of the three-step differentiation, significantly increased the differentiation efficiency. It also increased the glucose-stimulated insulin and C-peptide secretion in insulin-secreting cells derived from mES cells compared to the control green fluorescent protein (GFP) vector-transduced group. For the first time, we have demonstrated that the coexpression of Pdx1 and MafA during a specific time window of development can act synergistically with either Ngn3 or NeuroD to promote the differentiation of mES cells into insulin-secreting cells.
从胚胎干细胞（ES 细胞）或诱导多能干细胞（iPS 细胞）分化而来的胰腺 β 细胞在细胞治疗中是一种很有前途的策略。胰腺 β 细胞的发育受到转录因子分子网络的顺序表达调控。在本实验中，我们采用了三步分化方案，通过腺病毒载体在不同的分化时间以不同的组合过表达转录因子，将 mES（小鼠 ES）细胞分化为胰岛素分泌细胞。我们发现，Pdx1 和 MafA 与 Ngn3 或 NeuroD 的共表达，特别是在三步分化的最后阶段，显著提高了分化效率。与对照绿色荧光蛋白（GFP）载体转导组相比，它还增加了源自 mES 细胞的胰岛素分泌细胞中葡萄糖刺激的胰岛素和 C 肽分泌。我们首次证明，在发育的特定时间窗口内共表达 Pdx1 和 MafA 可以与 Ngn3 或 NeuroD 协同作用，促进 mES 细胞向胰岛素分泌细胞的分化。
