树鼩脐带间充质干细胞分离、培养及成骨成脂分化的鉴定  

Tree shrew umbilical cord mesenchymal stem cells: isolation, cultivation and osteogenic and adipogenic differentiation

作　　者：阮光萍[1] 朱路[1] 刘菊芬[1] 李自安[1] 王金祥[1] 庞荣清[1] 潘兴华[1] 

机构地区：[1]解放军成都军区昆明总医院细胞生物治疗中心,云南省昆明市650032

出　　处：《中国组织工程研究》2017年第9期1373-1377,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家科技支撑计划项目(2014BAI01B01);云南省科技计划项目(2013CA005)~~

摘　　要：背景:脐带间充质干细胞是组织工程研究中的理想种子细胞。目的:分离、培养及鉴定树鼩脐带间充质干细胞,建立标准化的树鼩脐带间充质干细胞株。方法:采用剖腹产分离树鼩脐带,用组织块贴壁法培养脐带间充质干细胞,用流式标记法检测脐带间充质干细胞表面标志,用成骨、成脂诱导培养基诱导脐带间充质干细胞向成骨细胞和脂肪细胞分化。结果与结论:(1)细胞鉴定:培养的脐带间充质干细胞CD90和CD105的阳性表达率分别为99.9%和99.8%,造血干细胞标志物CD34的表达率为0.0%,内皮细胞标志物CD31的表达率为0.7%,符合脐带间充质干细胞表面标志特征。(2)细胞形态:诱导后的脐带间充质干细胞茜素红染色观察到钙结节,油红O染色观察到脂滴。(3)结果证实,实验成功分离培养了具备成骨成脂分化能力的树鼩脐带间充质干细胞。BACKGROUND： Studies have shown that umbilical cord mesenchymal stem cells are ideal seed cells for tissue engineering research.OBJECTIVE：To isolate,culture and identify tree shrew umbilical cord mesenchymal stem cells,in order to establish a standardized tree shrew umbilical cord mesenchymal stem cell lines.METHODS：Caesarean-isolated tree shrew umbilical cord samples were used to isolate and culture umbilical cord mesenchymal stem cells using tissue explant adherent method.Flow cytometry assay was used to detect cell surface markers.Osteogenic and adipogenic induction media were used to induce umbilical cord mesenchymal stem cells to differentiate into osteoblasts and adipocytes.RESULTS AND CONCLUSION：The cultured umbilical cord mesenchymal stem cells expressed CD90 and CD105 with the positive rate of 99.9%and 99.8%respectively.Hematopoietic stem cell marker CD34 expression rate was 0.0%and the endothelial cell marker CD31 expression rate was 0.7%,in line with the characteristics of umbilical cord mesenchymal stem cell surface markers.Calcium nodules by alizarin red staining and lipid droplets by oil red O staining were observed in the induced cells.These experimental findings indicated that umbilical cord mesenchymal stem cells from tree shrews capable of osteogenic and adipogenic differentiation were successfully isolated and cultured.

关 键 词：干细胞 细胞分化 树鼩科 组织工程 脐带脐血干细胞 脐带间充质干细胞 树鼩 成骨分化 成脂分化 CD抗原 表面标志物 钙结节 脂滴 分离培养 种子细胞 

分 类 号：R722.12[医药卫生—儿科]