一种简单的人脐带间充质干细胞分离培养方法.PDFVIP

162 中华细胞与干细胞杂志(电子版) 2011 年 11 月第1卷第2期          Chin J Cell Stem Cell (Electronic Edition), Nov. 2011,Vol.1, No.2 •实验研究• 一种简单的人脐带间充质干细胞分离 培养方法 庞荣清  何洁  李福兵  赵晶  阮光萍  王桂华  瞿良  李黎  蔡学敏  潘兴华 【摘要】目的  建立一种简单的人脐带间充质干细胞分离培养方法。方法  取新鲜脐带， 剪成5 cm 长的小段，直接剪碎为糊状，加入含10 ﹪胎牛血清的DMEM/F12 在培养瓶中培养， 光学显微镜下观察细胞的生长特征，运用流式细胞仪检测分析细胞的抗原标志表达，并检测其 体外多向分化潜能。结果  运用不剥离血管组织、不用酶消化的组织贴块培养法可以从新鲜 脐带中分离到贴壁生长的成纤维细胞样细胞，这些细胞表达CD29、CD44、CD90 和CD105，并 可在体外诱导分化为脂肪、骨和软骨细胞。结论  运用不剥离血管组织、不用酶消化的组织贴 块培养法可以从人脐带中分离到间充质干细胞，其生物学特性与骨髓间充质干细胞相似。 【关键词】脐带间充质干细胞；分离；培养 A simple method for isolation of human umbilical cord mesenchymal stem cell PANG Rong- qing, HE Jie, LI Fu-bing, ZHAO Jing, RUAN Guang-ping, WANG Gui-hua, QU Liang, LI Li, CAI Xue-min, PAN Xing-hua. Center for Stem Cell and Tissue Organ Engineering, Kunming General Hospital, Kunming 650032, China 【Abstract 】Objective  To establish a simple method for isolation of human umbilical cord mesenchymal stem cell. Methods  Five-cm long segments from fresh umbilical cord were directly minced into pasty fine pieces, and the pieces were cultured in tissue flask with DMEM/F12 medium supplemented with 10 ﹪ fetal bovine serum. The morphological characteristic of the cells was investigated by light microscope, and their expression of mesenchymal surface markers were analyzed by flow cytometry. The multidifferentiation potential was examined in vitro, too. Results The fibroblast-like cells could be isolated from the fresh umbilical cord by a nonenzymatic isolation procedure without removing of the vessels. These adherent passage 3 cells expressed mesenchymal markers including CD29, CD44, CD90 and CD105, and also could be induced to differentiate into adipocytes, osteoblasts and chondrocytes. Conclussion  Mesenchymal stem cells, which biological characteristic was similar to bone marrow mesenchymal stem cells, could be isolated from fresh human umbilical co