密度梯度离心联合传代贴壁法分离、培养兔骨髓间充质干细胞

Isolation and culture rabbit BMMSCs by density gradient centrifugation combined with passage adherence method

目的探讨密度梯度离心联合传代贴壁法分离、培养兔骨髓间充质干细胞的可行性。方法应用密度梯度离心联合传代贴壁法分离、培养兔骨髓间充质干细胞,观察细胞形态变化,应用免疫荧光法鉴定细胞并计算纯度,应用八肽胆囊收缩素(CCK-8)法绘制细胞生长曲线。结果应用密度梯度离心联合传代贴壁法可以分离、培养兔骨髓间充质干细胞,第3代细胞接种后2~8 d进入细胞增殖对数期,兔骨髓间充质干细胞的纯度可达(81.77±2.10)%,3代细胞纯度组间比较差异有统计学意义(P<0.05)。结论应用密度梯度离心联合传代贴壁法可以分离、培养并获取正常活性及理想纯度的兔骨髓间充质干细胞。

Objective To explore the method of isolation and culture rabbit BMMSCs by density - gradients centrifuge and passages adherence cuhure. Methods The changes in cell morphology were observed, the BMMSCs was identified and the purity was calculated by immumofluorescence,and the growth curve of the cell was draw by CCK - 8 method. Results It was possible to isolate and culture rabbit BMMSCs by densitygradients centrifuge and passages adherence culture, the 3rd generation cells entered the cell＇s log period during the 2nd to 8th day after inoculation,the BMMSCs purity was （81.77 ± 2.10）% ,the difference between the three generations of cell purity were statistically significant （P 〈 0.05）. Conclusion It is possible to isolation and culture rabbit BMMSCs by density -gradients centrifuge and passages adherence culture method, the BMMSCs＇ activity and purity is desired.