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首页> 外文期刊>Journal of Cellular Physiology >Generation of Human Lens Epithelial-Like Cells From Patient-Specific Induced Pluripotent Stem Cells
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Generation of Human Lens Epithelial-Like Cells From Patient-Specific Induced Pluripotent Stem Cells

机译：一代的人工晶状体Epithelial-Like细胞从特定的诱导多功能干细胞细胞

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摘要

Cataractogenesis begins from the dynamic lens epithelial cells (LECs) and adjacent fiber cells. LECs derived from cell lines cannot maintain the crystalline expression as the primary LECs. The current study aimed to efficiently generate large numbers of human LECs from patient-specific induced pluripotent stem cells (iPSCs). Anterior lens capsules were collected from cataract surgery and were used to culture primary hLECs. iPSCs were induced from these primary hLECs by lentiviral transduction of Oct4, Sox2, Klf4, and c-Myc. Then, the generated iPSCs were re-differentiated into hLECs by the 3-step addition of defined factor combinations (Noggin, BMP4/7, bFGF, and EGF) modified from an established method. During the re-differentiation process, colonies of interest were isolated using a glass picking tool and cloning cylinders based on the colony morphology. After two steps of isolation, populations of LEC-like cells (LLCs) were generated and identified by the expression of lens marker genes by qPCR, western blot and immunofluorescence staining. The study introduced a modified protocol to isolate LLCs from iPSCs by defined factors in a short time frame. This technique could be useful for mechanistic studies of lens-related diseases. (C) 2016 Wiley Periodicals, Inc.
机译：Cataractogenesis从动态镜头开始上皮细胞(lec)和相邻纤维细胞。lec源自细胞系无法维持水晶表达式作为主要的lec。本研究旨在有效地生成人类lec从特定病人的数量诱导多能干细胞(万能)。镜头胶囊收集从白内障手术和用于主hLECs文化。从这些主要hLECs则被诱导慢病毒转导Oct4、Sox2 Klf4,原癌基因。re-differentiated hLECs的3步骤添加定义因素组合(‘诺金’,BMP4/7、bFGF和EGF)的修改建立方法。感兴趣的过程中,殖民地是孤立的使用一个玻璃选择工具和基于克隆气缸菌落形态。隔离,LEC-like细胞的数量(有限责任公司)生成和确定的表达吗的镜头qPCR标记基因,蛋白质印迹免疫荧光染色。修改后的协议由万能干细胞隔离llc在短时间内定义的因素。技术可以用于机械的研究lens-related疾病。期刊、公司。

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