4℃保存条件下存放时间对大鼠嗅球来源的嗅鞘细胞活力的影响

The influence of 4 degree centigrade concervation on cells activity of rats' olfactory bulbs derived olfactory ensheathing cells

目的观察大鼠嗅鞘细胞4℃保存下不同时间对细胞活力的影响,为临床细胞移植治疗的细胞标准化制备提供依据。方法无菌取7d龄大鼠嗅球,胰蛋白酶消化培养,2～3d后计数,D/F12悬浮,调细胞终浓度为2000/μl,置于4℃冰箱,分别在不同时间依次取出,每管分别行台盼蓝染色,噻唑兰(MTT)法实验,P75免疫荧光染色,检测其活力变化。结果 4℃保存2h以内细胞活力达85%,超过3h细胞活力显著下降,免疫组化显示细胞状态也有明显改变,由规则的梭形或三角形变成不规则的多态形。结论 4℃2h为临床应用前细胞存放的标准时间规范,临床必需遵循。

Objective To detect the influence of 4 degree centigrade concervation on cells activity of rat olfactory ensheathing cells to provide evidences for clinical transplantation. Methods 7 days rat olfactory bulbs were taken out sterilitelly, digested with parenzyme and then cultivated and counted for 2 to 3 days. The cells were floated in D/F12 and cells concentration was adjusted to 106/50μl per tube, then the cells suspension was conserved in refrigerator of 4 degree centigrade. The tubes were taken out one by one in different times, stained with trypanblue for every tube and MTT method respectively, p75 immunofluorescence was used to detect the activity changes. Results The activity of the cells that conserved in 4℃ for less than two hours can still reach about 85 percent. However, the cells activity decreased significantly and the cells’ condition changed obviously from regular fusiform into irregular multi-shapes while conserved for more than 3 hours. Conclusions Cells conserved in 4℃ for less than 2h can meet the transplantation need.