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Potential Factors for the Differentiation of ESCs/iPSCs into Insulin-Producing Cells

机译：ESCs / iPSCs分化为胰岛素产生细胞的潜在因素

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The low efficiency of in vitro differentiation of human embryonic stem cells (ESCs) or human induced pluripotent stem cells (iPSCs) into insulin-producing cells thus creates a crucial hurdle for the clinical implementation of human pluripotent stem cells (PSCs). In this study, we investigated the key factors for the differentiation of PSCs into insulin-producing cells. We obtained microarray data of HUES8 and HUES6 from two GeneChips (GPL3921: Affymetrix HT Human Genome U133A Array, GPL570: Affymetrix Human Genome U133 Plus 2.0 Array) in a database of GEO (NCBI), since HUES8 can differentiate into pancreatic cells, while HUES6 hardly demonstrates any differentiation at all. The genes with more than fourfold higher expressions in HUES8 compared to HUES6 included RPS4Y1, DDX3Y, EIF1AY, GREM1, GATA6, and NLGN4Y. Since there were four genes, RPS4Y1, DDX3Y, EIF1AY, and NLGN4Y, on the Y chromosome and HUES8 was a male cell line and HUES6 was a female cell line, we excluded these genes in this study. On the other hand, genes with more than fourfold higher expressions in HUES6 compared to HUES8 included NLRP2, EGR1, and SMC3. We next compared iPSCs derived from pancreatic cells (PiPSCs) and iPSCs derived from fibroblasts (FiPSCs). PiPSCs differentiated into insulin-producing cells more easily than FiPSCs because of their epigenetic memory. The gene expressions of GREM1, GATA6, NLRP2, EGR1, and SMC3 in PiPSCs and FiPSCs were also investigated. The expression level of GREM1 and GATA6 in PiPSCs were higher than in FiPSCs. On the other hand, EGR1, which was lower in HUES8 than in HUES6, was predictably lower in PiPSCs than FiPSCs, while NLRP2 and SMC3 were higher in PiPSCs than FiPSCs. These data suggest that the expression of GATA6 and GREM1 and the inhibition of EGR1 may be important factors for the differentiation of PSCs into insulin-producing cells.
机译：人胚胎干细胞（ESC）或人诱导的多能干细胞（iPSC）体外分化为产生胰岛素的细胞的效率低下，因此为人多能干细胞（PSC）的临床实施创造了关键的障碍。在这项研究中，我们调查了PSCs分化为产生胰岛素的细胞的关键因素。我们从GEO（NCBI）数据库中的两个GeneChips（GPL3921：Affymetrix HT人类基因组U133A阵列，GPL570：Affymetrix人类基因组U133 Plus 2.0阵列）中获得了HUES8和HUES6的微阵列数据，因为HUES8可以分化为胰腺细胞，而HUES6几乎没有任何区别。与HUES6相比，HUES8中具有高四倍以上表达的基因包括RPS4Y1，DDX3Y，EIF1AY，GREM1，GATA6和NLGN4Y。由于Y染色体上有RPS4Y1，DDX3Y，EIF1AY和NLGN4Y这四个基因，HUES8是雄性细胞系，HUES6是雌性细胞系，因此在本研究中我们将这些基因排除在外。另一方面，与HUES8相比，HUES6中表达高四倍以上的基因包括NLRP2，EGR1和SMC3。接下来，我们比较了源自胰腺细胞的iPSC（PiPSC）和源自成纤维细胞的iPSC（FiPSC）。由于它们的表观遗传记忆，PiPSC比FiPSC更容易分化为产生胰岛素的细胞。还研究了PiPSC和FiPSC中GREM1，GATA6，NLRP2，EGR1和SMC3的基因表达。 PiPSC中GREM1和GATA6的表达水平高于FiPSC。另一方面，PiPSC中的EGR1在HUES8中低于HUES6，在PiPSC中可预测地低于FiPSC，而NLRP2和SMC3在PiPSC中则高于FiPSC。这些数据表明，GATA6和GREM1的表达以及对EGR1的抑制可能是将PSC分化为胰岛素产生细胞的重要因素。

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