1 Bioresource Collection and Research Center, Food Industry Research and Development Institute , Hsinchu, Taiwan .
Stem Cells Dev. 2014 Feb 15;23(4):372-9. doi: 10.1089/scd.2013.0253. Epub 2013 Nov 16.
Human embryonic stem cells (hESCs), due to their self-renewal capacity and pluripotency, have become a potential source of transplantable β-cells for the treatment of diabetes. However, it is imperative that the derived cells fulfill the criteria for clinical treatment. In this study, we replaced common Matrigel with a synthetic peptide-acrylate surface (Synthemax) to expand undifferentiated hESCs and direct their differentiation in a defined and serum-free medium. We confirmed that the cells still expressed pluripotent markers, had the ability to differentiate into three germ layers, and maintained a normal karyotype after 10 passages of subculture. Next, we reported an efficient protocol for deriving nearly 86% definitive endoderm cells from hESCs under serum-free conditions. Moreover, we were able to obtain insulin-producing cells within 21 days following a simple three-step protocol. The results of immunocytochemical and quantitative gene expression analysis showed that the efficiency of induction was not significantly different between the Synthemax surface and the Matrigel-coated surface. Thus, we provided a totally defined condition from hESC culture to insulin-producing cell differentiation, and the derived cells could be a therapeutic resource for diabetic patients in the future.
人胚胎干细胞（hESCs）由于其自我更新能力和多能性，已成为治疗糖尿病的可移植β细胞的潜在来源。然而，衍生细胞必须满足临床治疗的标准。在这项研究中，我们用合成肽-丙烯酸盐表面（Synthemax）代替普通的 Matrigel 来扩增未分化的 hESCs，并在定义的无血清培养基中指导其分化。我们证实，细胞仍然表达多能标记物，具有分化为三个胚层的能力，并在传代培养 10 代后保持正常核型。接下来，我们报告了一个有效的方案，即在无血清条件下从 hESC 中获得近 86%的确定内胚层细胞。此外，我们能够在简单的三步方案后 21 天获得产生胰岛素的细胞。免疫细胞化学和定量基因表达分析的结果表明，在 Synthemax 表面和 Matrigel 涂层表面之间，诱导效率没有显著差异。因此，我们从 hESC 培养到产生胰岛素的细胞分化提供了一个完全定义的条件，衍生细胞将来可能成为糖尿病患者的治疗资源。
