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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 observe the effect of different time on the viability of rat olfactory ensheathing cells (NSCs) preserved at 4 ℃, and provide the basis for the standard preparation of cells for clinical cell transplantation. Methods The olfactory bulb of 7-day-old rats were aseptically cultured and digested with trypsin. The cells were counted after 2 to 3 days and suspended in D / F12. The final cell concentration was adjusted to 2000 / μl. The cells were placed in 4 ℃ refrigerator and taken out at different times. Trypan blue staining, MTT assay and P75 immunofluorescence staining were used to detect the changes of viability. Results The viability of cells reached 85% within 2 hours after preservation at 4 ° C. The viability of cells over 3 hours significantly decreased. The immunohistochemistry showed that the cellular status also changed significantly from regular fusiform or triangular to irregular polymorphism. Conclusions 4 ℃ 2h is the standard time standard for storing cells before clinical application, so clinical follow-up is needed.