子宫结合带干细胞的培养与鉴定

目的建立子宫结合带干细胞（uJZSCs）的分离培养及鉴定方法。方法将手术中无菌取得的子宫内肌层即子宫结合带用胰酶和胶原酶Ⅰ进行消化、培养、扩增。采用倒置显微镜观察细胞形态,流式细胞仪进行表型分析及存活率判断,CCK-8法判断其生长特性,并在体外诱导其成脂、成骨、成软骨分化。结果所获细胞传代后能迅速贴壁,呈长梭形;流式细胞仪检测结果显示,表面标志物CD90、CD73、CD105、CD29、CD44、CD13、CD166、HLAABC呈阳性表达,而CD34、CD45、CD14、HLA-DR、CD19呈阴性表达。CCK-8生长曲线呈S形。诱导分化实验结果显示,uJZSCs具有成脂、成骨和成软骨的分化...

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Published in解放军医学杂志 Vol. 40; no. 3; pp. 173 - 177
Main Author 康康 王蔼明 尹善德 赵勇 王明凯 黄绍敏
Format Journal Article
LanguageChinese
Published 2015
Subjects
子宫结合带
表型
间质干细胞
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ISSN0577-7402

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Summary:目的建立子宫结合带干细胞（uJZSCs）的分离培养及鉴定方法。方法将手术中无菌取得的子宫内肌层即子宫结合带用胰酶和胶原酶Ⅰ进行消化、培养、扩增。采用倒置显微镜观察细胞形态,流式细胞仪进行表型分析及存活率判断,CCK-8法判断其生长特性,并在体外诱导其成脂、成骨、成软骨分化。结果所获细胞传代后能迅速贴壁,呈长梭形;流式细胞仪检测结果显示,表面标志物CD90、CD73、CD105、CD29、CD44、CD13、CD166、HLAABC呈阳性表达,而CD34、CD45、CD14、HLA-DR、CD19呈阴性表达。CCK-8生长曲线呈S形。诱导分化实验结果显示,uJZSCs具有成脂、成骨和成软骨的分化能力。结论 uJZSCs具有很强的增殖扩增能力,可作为间充质干细胞的来源。
Bibliography:mesenchymal stem cells; uterine junctional zone; phenotype
11-1056/R
Objective To establish a method for isolation, culture, and identification of stem cells in uterine junctional zone （uJZSCs）. Methods Specimens of uterus muscle layer at uterine junctional zone （uJZ） were harvested under aseptic condition, and they digested, cultivated and amplified using trypsogen and collagenase I. The morphology of uJZSCs was observed with inverted microscope, and cell viability and phenotype were analyzed by flow cytometry. The adipogenic, osteogenic and chondrogenic differentiation was induced in vitro, and their biological growth characteristic was identified by CCK-8 marking. Results The cells were adhered after passage, and they presented long spindle in shape. Flow cytometry showed that the expressions of CD90, CD73, CD105, CD29, CD44, CD13, CD166, and HLA-ABC marking was also positive, while they were negative for CD34, CD45, CD14, HLA-DR and CD19. The CCK-8 growth curve was S-shaped. The induced differentiation expe
ISSN:0577-7402