山羊骨髓间充质干细胞的分离培养及鉴定

Isolation culture and characterization of bone marrow mesenchymal stem cell from goat

目的分离培养山羊的间充质干细胞,并为其在骨组织工程研究中的应用奠定基础。方法应用常规的密度梯度离心法结合贴壁培养法从山羊骨髓中分离间充质干细胞,并通过观察分离培养细胞的形态,检测其表面的CD分子表达,以及RTPCR和免疫组化病理染色方法证实成骨、成软骨、成脂肪多方向分化潜能来对细胞进行鉴定。结果分离的细胞具有成纤维细胞样形态,第三代细胞的表面分子CD29和CD44阳性率为98.70%、99.54%,而CD62L和CD45阳性率为1.10%和0.73%。RT-PCR和免疫组化病理染色证实分离细胞具有成骨、成软骨、成脂肪多方向分化潜能。结论成功地分离培养山羊的骨髓间充质干细胞,使山羊间充质干细胞应用于骨组织工程的研究成为可能。

Objective To isolate and culture goat mesenchymal stem cells （MSCs） from bone marrow, and explore their application in bone tissue engineering. Methods Cells were isolated from goat bone marrow by conventional methods. The identification of cell was made by cell morphous, expression of cell epitope profile, and multilineage differentiation of osteogenesis, chondrogenesis, and adipogenesis assayed RT-PCR and immunohistology staining. Results The cells were fibroblast-like, whose positive rate of CD29 and CD44 were 98.70% and 99.54% ,and the percentage of CD62L and CD45 were 1. 10% and 0.73%. The results of RT-PCR and immunohistology staining showed muhilineage differentiation potential of cell. Conclusion MSCs were successfully isolated from goat bone marrow and it is possible that they will be used in the research field of bone tissue engineering.