基础研究子宫结合带干细胞的培养与鉴定.PDF

Med J Chin PLA, Vol. 40, No. 3, March 1, 2015 　173 论　著 ·基础研究· 子宫结合带干细胞的培养与鉴定 康康，王蔼明，尹善德，赵勇，王明凯，黄绍敏 [摘要]　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 、HLA- ABC 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。 [关键词]　间质干细胞；子宫结合带；表型 [中图分类号]　R329.24 ；R394.26　　　　[文献标志码]　A　　　　[文章编号]　0577-7402(2015)03-0173-05 [DOI]　10.11855/j.issn.0577-7402.2015.03.01 Culture and identification of stem cells of uterine junctional zone KANG Kang1 1,2* 2 2 2 1 , WANG Ai-ming , YIN Shan-de , ZHAO Yong , WANG Ming-kai , HUANG Shao-min 1 Medical School of Chinese PLA, Beijing 100853, China 2Department of Obstetrics and Gynecology, Navy General Hospital of PLA, Beijing 100048, China * Corresponding author, E-mail: one-army@ This work was s upported by the K ey P rojects of “Twelfth Five-Year P lan” N ational Science Technology P illar P rogram (2012BAI32B05); the National Natural Science Foundation of China ; and the Project of M ilitary L ogistics Scientific R esearch (CHJ13J012) [Abstract]　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 usi ng trypsogen and collagenase Ⅰ. The morphology of uJZS