罗曼鹤鸡骨髓间充质干细胞的分离培养和鉴定

目的建立鸡骨髓间充质干细胞（BMSCs）的分离培养和鉴定体系。方法从1～14天龄罗曼鹤鸡骨髓中分离骨髓细胞，差速贴壁法纯化、扩增BMSCs。倒置显微镜下观察细胞形态特征，MTY法绘制生长曲线，流式细胞仪检测细胞标志物，并分别进行成骨、成脂诱导分化能力检测。结果分离培养的细胞呈成纤维细胞样或长梭形，生长状态良好；CD29阳性表达率为92．10％，CD34阳性表达率仅为0．80％；经成骨诱导分化，出现明显的钙化结节，茜素红染色阳性；经成脂诱导分化，油红O染色阳性，细胞内出现明显的脂质小滴。结论建立了操作简单、高效的鸡BMSCs分离培养和鉴定体系，为鸡BMSCs的进一步研究和应用提供了良好的基础。

Isolation,cultivation and identification of chicken bone marrow mesenchymal stem cells

Objective To establish the isolation-cultivation-identification system of chicken bone marrow mesenchymal stem cells （BMSCs）. Methods Bone marrow cells were isolated from 1-14 days old chickens. Then, BMSCs were puri- fied and cultured by the whole bone marrow adherent method. Morphological characteristics were observed by inverted mi- croscope, growth curve were found by MTY, and flow cytometry was used to detect the surface markers. The osteoblast and adipocyte differentiation was induced. Results The cultured BMSCs were fibroblast-like or long spindle with good growth condition. The positive rate of CD29 was 92.10% and the positive rate of CD34 was only 0.80%. After being induced to dif- ferentiate osteoblast, they showed obvious calcification nodules, and alizarin red staining was positive; after the adipogenic differentiation, oil red O staining was positive, and lipid droplets appeared in the cells. Conclusion The isolation-cultiva- tion-identification system of chicken BMSCs with simple operation and high efficiency is successfully established, which al- so provides better basis for further research of chicken BMSCs.