Aberdam Edith, Petit Isabelle, Sangari Linda, Aberdam Daniel
INSERM U976 and Université Paris-Diderot, Hôpital St-Louis, Paris, France.
PLoS One. 2017 Jun 22;12(6):e0179913. doi: 10.1371/journal.pone.0179913. eCollection 2017.
Induced pluripotent stem cells hold great potential to produce unlimited amount of differentiated cells as cellular source for regenerative medicine but also for in vitro drug screening and cytotoxicity tests. Ocular toxicity testing is mandatory to evaluate the risks of drugs and cosmetic products before their application to human patients by preventing eye irritation or insult. Since the global ban to use animals, many human-derived alternatives have been proposed, from ex-vivo enucleated postmortem cornea, primary corneal cell culture and immortalized corneal epithelial cell lines. All of them share limitations for their routine use. Using an improved protocol, we derived limbal epithelial cells from human induced pluripotent stem cells, named LiPSC, that are able to be passaged and differentiate further into corneal epithelial cells. Comparative RT-qPCR, immunofluorescence staining, flow cytometry analysis and zymography assays demonstrate that LiPSC are morphologically and molecularly similar to the adult stem cells. Moreover, contrary to HCE, LiPSC and primary limbal cells display similarly sensitive to cytotoxicity treatment among passages. Our data strongly suggest that LiPSC could become a powerful alternative cellular model for cosmetic and drug tests.
诱导多能干细胞具有巨大潜力，可产生无限量的分化细胞，作为再生医学的细胞来源，也可用于体外药物筛选和细胞毒性测试。眼部毒性测试对于评估药物和化妆品在应用于人类患者之前的风险至关重要，可预防眼部刺激或损伤。自从全球禁止使用动物以来，已经提出了许多源自人类的替代方法，包括离体摘除的死后角膜、原代角膜细胞培养和永生化角膜上皮细胞系。它们在常规使用中都存在局限性。我们采用改进的方案，从人诱导多能干细胞中获得了角膜缘上皮细胞，命名为LiPSC，这些细胞能够传代并进一步分化为角膜上皮细胞。比较逆转录定量聚合酶链反应、免疫荧光染色、流式细胞术分析和酶谱分析表明，LiPSC在形态和分子水平上与成体干细胞相似。此外，与角膜内皮细胞相反，LiPSC和原代角膜缘细胞在传代过程中对细胞毒性处理表现出相似的敏感性。我们的数据强烈表明，LiPSC可能成为化妆品和药物测试的强大替代细胞模型。
