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人胚胎干细胞和诱导多能干细胞源性角膜内皮细胞的比较转录组分析。

Comparative transcriptome analysis of hESC- and iPSC-derived corneal endothelial cells.

机构信息

The Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, 21287, United States.

The Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, 21287, United States.

出版信息

Exp Eye Res. 2018 Nov;176:252-257. doi: 10.1016/j.exer.2018.08.023. Epub 2018 Sep 6.

Abstract

The corneal endothelium (CE), a monolayer of hexagonal cells constitutes the innermost layer of the cornea that is critical in maintaining clarity by mediating hydration through barrier and pump functions. Corneal endothelial cells (CECs) have limited proliferative potential and therefore generation of CECs has been undertaken by many groups. We previously reported generation of CECs from peripheral blood mononuclear cell (PBMC)-originated, induced pluripotent stem cells (iPSCs). In here, we extend our analysis through next-generation seqeuncing based transcriptome profiling of H9 human embryonic stem cell (hESC)- and human PBMC-originated, iPSC-derived CECs. The differentiating CECs on day 20 (D20) exhibited a tightly packed hexagonal/polygonal shape expressing zona occludens-1 (ZO-1) and N-cadherin at the cell boundaries. Next-generation RNA sequencing of hESC- and iPSC-derived CECs detected expression (≥0.659 RPKM) of 13,546 and 13,536 genes, respectively. Comparative transcriptome analysis of hESC- and iPSC-derived CECs revealed 13,208 (>96%) genes common in both transcriptomes. Among the 13,208 genes common in these transcriptomes, 12,580 (>95%) exhibited a quantitatively similar expression. To the best of our knowledge, this is the first report presenting comparative transcriptome analysis of hESC- and iPSC-derived CECs.

摘要

角膜内皮(CE)是一层六边形细胞,构成了角膜的最内层,通过屏障和泵功能介导水合作用,对保持角膜的透明度至关重要。角膜内皮细胞(CEC)的增殖能力有限,因此许多研究小组已经开展了 CEC 的生成工作。我们之前曾报道过从外周血单核细胞(PBMC)来源的诱导多能干细胞(iPSC)生成 CEC。在这里,我们通过下一代测序(基于转录组分析)对 H9 人胚胎干细胞(hESC)和人 PBMC 来源的、iPSC 衍生的 CEC 进行了扩展分析。第 20 天(D20)的分化 CEC 呈紧密包装的六边形/多边形形状,在细胞边界表达封闭蛋白-1(ZO-1)和 N-钙粘蛋白。hESC 和 iPSC 衍生的 CEC 的下一代 RNA 测序分别检测到 13546 和 13536 个基因的表达(≥0.659 RPKM)。hESC 和 iPSC 衍生的 CEC 的比较转录组分析显示,这两个转录组中有 13208 个(>96%)基因是共同的。在这两个转录组中共有的 13208 个基因中,有 12580 个(>95%)表现出定量相似的表达。据我们所知,这是首次报道 hESC 和 iPSC 衍生的 CEC 比较转录组分析。

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