【摘要】 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,MTT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。更多还原

【Abstract】 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 purified and cultured by the whole bone marrow adherent method. Morphological characteristics were observed by inverted microscope,growth curve were found by MTT,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 CD29was 92. 10% and the positive rate of CD34was only 0. 80%. After being induced to differentiate 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-cultivation-identification system of chicken BMSCs with simple operation and high efficiency is successfully established,which also provides better basis for further research of chicken BMSCs.更多还原