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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 relates to a method for inducing differentiation of inducible pluripotent stem cells to cultivate corneal epithelial cells, which method comprises culturing the corneal epithelial cells containing DMEM F12, L-ascorbic acid, sodium selenite, Preparing a basic medium; Adding an inducer for culturing an inducible pluripotent stem cell to the basal medium; Preparing a feeder-free culture medium by coating the basic medium with the vitronectin recombinant human protein; Adding the inducer pluripotent stem cells to the feeder-containing medium; Culturing the induced pluripotent stem cells cultured in the feeder-containing medium in a differentiation medium; Adding the BMP4 to the differentiation medium to differentiate the cultured induced pluripotent stem cells into corneal stem cells; A step of sequentially adding Wnt3a and miRNA or differentiation of Wnt3a and miRNA to differentiate into corneal epithelial cells after the step of adding BMP4 to the differentiation medium; And culturing the cells differentiated from the induced pluripotent stem cells in a PI culture medium.
机译:本发明涉及一种诱导可诱导的多能干细胞分化以培养角膜上皮细胞的方法,该方法包括培养含有DMEM F12,L-抗坏血酸,亚硒酸钠的角膜上皮细胞,制备碱性培养基。向基础培养基中添加用于培养可诱导的多能干细胞的诱导剂;通过用玻连蛋白重组人蛋白包被基础培养基来制备无饲养层的培养基;将诱导剂多能干细胞添加到含饲养层的培养基中;在分化培养基中培养在含饲养层的培养基中培养的诱导多能干细胞;将BMP4加入分化培养基中,以将培养的诱导多能干细胞分化为角膜干细胞。在向分化培养基中添加BMP4的步骤之后,依次添加Wnt3a和miRNA或分化Wnt3a和miRNA以分化为角膜上皮细胞的步骤;然后在PI培养基中培养从诱导的多能干细胞中分化出来的细胞。

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