ROCK Inhibition Extends Passage of Pluripotent Stem Cell-Derived Retinal Pigmented Epithelium
ROCK Inhibition Extends Passage of Pluripotent Stem Cell-Derived Retinal Pigmented Epithelium
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DOI：
10.5966/sctm.2014-0079
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发表时间：
2014-09-01
影响因子：
6
通讯作者：
Clegg, Dennis O.
中科院分区：
文献类型：
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作者：
Croze, Roxanne H.;Buchholz, David E.;Clegg, Dennis O.
人类胚胎干细胞（hESC）为新兴的基于细胞的疗法提供了潜在的无限细胞供应。不幸的是，获得不同细胞类型的过程可能是耗时且昂贵的。在发达国家，年龄相关性黄斑变性（AMD）是老年人失明的主要原因，仅在美国就有720多万人受到影响。hESC衍生的视网膜色素上皮细胞（hESC-RPE）和诱导多能干细胞衍生的RPE（iPSC-RPE）都正在被多个小组开发用于AMD治疗，但它们在培养中扩增的潜力有限。为了试图克服这种传代限制，我们检查了Rho相关的卷曲螺旋蛋白激酶（ROCK）在hESC-RPE和iPSC-RPE培养物中的参与。我们报道用Y-27632抑制ROCK 1/2允许hESC-RPE和iPSC-RPE的延长传代。微阵列分析表明，ROCK抑制可能通过多种途径抑制上皮细胞向间充质细胞的转化。这些包括抑制转化生长因子-β途径（TGFB 1和GDF 6）和Wnt信号传导的关键配体。两个重要的过程受到影响，允许hESC-RPE扩增增加。首先，ROCK抑制通过诱导参与细胞周期进程的多种组分来促进增殖。第二，ROCK抑制作用影响许多可能会聚以抑制RPE向间充质转化的途径。这允许hESC-RPE在培养物中保持功能延长但有限的时间。
Human embryonic stem cells (hESCs) offer a potentially unlimited supply of cells for emerging cell-based therapies. Unfortunately, the process of deriving distinct cell types can be time consuming and expensive. In the developed world, age-related macular degeneration (AMD) is the leading cause of blindness in the elderly, with more than 7.2 million people afflicted in the U.S. alone. Both hESC-derived retinal pigmented epithelium (hESC-RPE) and induced pluripotent stem cell-derived RPE (iPSC-RPE) are being developed for AMD therapies by multiple groups, but their potential for expansion in culture is limited. To attempt to overcome this passage limitation, we examined the involvement of Rho-associated, coiled-coil protein kinase (ROCK) in hESC-RPE and iPSC-RPE culture. We report that inhibiting ROCK1/2 with Y-27632 allows extended passage of hESC-RPE and iPSC-RPE. Microarray analysis suggests that ROCK inhibition could be suppressing an epithelial-to-mesenchymal transition through various pathways. These include inhibition of key ligands of the transforming growth factor-beta pathway (TGFB1 and GDF6) and Wnt signaling. Two important processes are affected, allowing for an increase in hESC-RPE expansion. First, ROCK inhibition promotes proliferation by inducing multiple components that are involved in cell cycle progression. Second, ROCK inhibition affects many pathways that could be converging to suppress RPE-to-mesenchymal transition. This allows hESC-RPE to remain functional for an extended but finite period in culture.