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Method for enriching highly functional differentiation-inducing cells and population of highly functional differentiation-inducing cells

机译:用于富集高功能分化诱导细胞和高官能分化诱导细胞的方法

摘要

To provide methods for selecting and concentrating highly functional differentiation-induced cells from a pluripotent stem cell-derived differentiation-induced cell group, and additionally to provide a highly functional differentiation-induced cell population concentrated by the concentration method.SOLUTION: The present invention provides a method by introducing a drug resistance gene into the gene locus of drug metabolizing enzyme CYP of pluripotent stem cells. Highly functional differentiation-induced cells, specifically, CYP high expression differentiation-induced hepatocytes and CYP high expression differentiation-induced small intestinal epithelial cells can be selected and concentrated by subjecting pluripotent stem cells in which the drug resistance gene is induced to differentiation-inducing treatment and by culturing obtained differentiation-induced cells in the presence of the drug corresponding to the drug resistance. It was confirmed that a highly functional differentiation-induced hepatocyte population obtained by concentration expresses CYP3A4, CYP1A2, CYP2C19, and the like which are markers of mature hepatocytes, equally to human primary culture hepatocytes, and a highly functional differentiation-induced small intestinal epithelial cell population expresses Villin, ISX, ANPEP, and the like which are small intestinal epithelial cell markers, equally to human small intestinal cells.SELECTED DRAWING: Figure 21
机译:提供从多能干细胞衍生的分化诱导的细胞基团中选择和浓缩高官能分化诱导的细胞的方法,并且另外提供由浓缩法浓缩的高官能分化诱导的细胞群。溶液:本发明提供了本发明通过将药物抗性基因引入多能干细胞药物代谢酶CYP的基因位点来实现方法。具有高稳定性分化诱导的细胞,具体而言,Cyp高表达分化诱导的肝细胞和CYP高表达分化诱导的小肠上皮细胞可以通过对受试者进行多能干细胞来选择和浓缩,其中诱导药物抗性基因对分化诱导治疗并且通过在对应于耐药性的药物存在下培养得到的分化诱导的细胞。证实,通过浓缩的浓度表达CYP3A4,CYP1A2,CYP2C19等具有成熟肝细胞的标记,同等为人初级培养肝细胞,以及高官能分化诱导的小肠上皮细胞人口表达villin,ISX,ANPEP等,这是小肠上皮细胞标志物,同等为人类的小肠细胞。选择图:图21

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