树鼩脐带间充质干细胞分离、培养及成骨成脂分化的鉴定

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 tissueengineering research.OBJECTIVE: To isolate, culture and identify tree shrew umbilical cord mesenchymal stem cells, in order toestablish 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 cordmesenchymal stem cells using tissue explant adherent method. Flow cytometry assay was used to detect cellsurface markers. Osteogenic and adipogenic induction media were used to induce umbilical cord mesenchymalstem 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 ratewas 0.0% and the endothelial cell marker CD31 expression rate was 0.7%, in line with the characteristics of umbilicalcord mesenchymal stem cell surface markers. Calcium nodules by alizarin red staining and lipid droplets by oil red Ostaining were observed in the induced cells. These experimental findings indicated that umbilical cord mesenchymalstem cells from tree shrews capable of osteogenic and adipogenic differentiation were successfully isolated and cultured.