云南省自然基金项目(2011HB050,2013DA004)。
目的 建立食蟹猴脐带间充质干细胞的分离培养方法。方法 新鲜食蟹猴脐带剪碎为糊状, 用含10%胎牛血清的DMEM/F12培养液培养, 观察细胞形态特征, 流式细胞技术分析细胞抗原标志的表达, 并检测其体外多向分化潜能。结果 运用组织贴块法可以从新鲜脐带中分离到贴壁生长、阳性表达CD29、CD44、CD90的成纤维细胞样细胞。这些细胞在体外诱导培养后可分别检测到脂滴、骨和软骨细胞。结论 运用组织贴块培养法可用含10%胎牛血清的DMEM/F12培养液从食蟹猴脐带中分离到间充质干细胞。
Objective To establish a method for isolation of cynomolgus monkey umbilical cord mesenchymal stem cells.Methods Fresh cynomolgus monkey umbilical cord was 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 characteristics of the resulting cells were examined, and their expression of mesenchymal cell surface markers were analyzed by flow cytometry.The multidifferentiation potential was examined in vitro, too.Results The fibroblast-like cells were successfully isolated from the fresh umbilical cord by an adherent culture procedure.These adherent cells expressed mesenchymal markers including CD29, CD44, and CD90, and also could be induced to differentiate into adipocytes, osteoblasts and chondrocytes.Conclusion Mesenchymal stem cells can be isolated from fresh cynomolgus monkey umbilical cord by using an adherent culture procedure.
庞荣清,何洁,李瑞生,赵晶,朱慧,朱向情,阮光萍,潘兴华.食蟹猴脐带间充质干细胞的分离与鉴定[J].中国比较医学杂志,2015,25(4):66~69.
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