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miRNA Method for Culturing Cornea Epithelial Cell by Inducing Differentiation of Induced Pluripotent Stem Cell Using miRNA and System for the Same

机译:通过使用miRNA诱导多能干细胞分化诱导miRNA培养角膜上皮细胞的方法

摘要

The present invention provides a method for culturing corneal epithelial cells by inducing differentiation of induced pluripotent stem cells. More specifically, the method comprises the following processes of: preparing a basal medium containing DMEM F12, L-ascorbic acid, sodium selenite, and sodium chloride; adding an additive for culturing induced pluripotent stem cells to the basal medium; coating the basal medium with vitronectin recombinant human proteins to prepare a feeder-free culture medium; adding the induced pluripotent stem cells to the feeder-free culture medium to culture the same; culturing the induced pluripotent stem cells cultured in the feeder-free culture medium, in a differentiation culture medium; adding BMP4 to the differentiation culture medium in order to differentiate the cultured induced pluripotent stem cells into corneal epithelial cells; after adding the BMP4 to the differentiation culture medium, in order to differentiate the induced pluripotent stem cells into corneal epithelial cells, adding Wnt3a and miRNA successively or simultaneously; and culturing the cells differentiated from the induced pluripotent stem cells in a PI culture fluid.
机译:本发明提供了通过诱导诱导的多能干细胞分化来培养角膜上皮细胞的方法。更具体地,该方法包括以下过程:制备包含DMEM F12,L-抗坏血酸,亚硒酸钠和氯化钠的基础培养基;向基础培养基中添加用于培养诱导多能干细胞的添加剂;用玻连蛋白重组人蛋白包被基础培养基以制备无饲养层的培养基;将诱导的多能干细胞添加到无饲养层的培养基中进行培养;在分化培养基中培养在无饲养层培养基中培养的诱导多能干细胞;向分化培养基中添加BMP4,以将培养的诱导多能干细胞分化为角膜上皮细胞。向分化培养基中加入BMP4后,为了将诱导的多能干细胞分化为角膜上皮细胞,先后或同时加入Wnt3a和miRNA。在PI培养液中培养从诱导的多能干细胞分化而来的细胞。

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