新生SD大鼠坐骨神经雪旺细胞培养与纯化的方法学研究

An experimental study of purifying cultured Schwann cells from newly born SD rats

目的探讨简单、可靠地获取大量纯度高、活力强的雪旺细胞的细胞培养方法。方法选用新生4～6d的SD大鼠,解剖双侧坐骨神经,在解剖镜下剥离去除神经外膜,获得神经束。将其剪碎,采用双酶二次消化法消化,DMEM培养液培养4d后应用G-418纯化,4d后对培养的细胞进行细胞计数、活力测定和免疫细胞化学鉴定。结果该法从新生大鼠双侧坐骨神经可提取3.62×106个雪旺细胞,纯化后经S-100蛋白免疫细胞化学染色,雪旺细胞的纯度可达96%以上。细胞活力强,经培养8d后即可进行传代。结论该方法可以获得纯度高、活力强的大量雪旺细胞,满足组织工程人工神经的需要。

Objective To introduce a simple way to obtain Schwann cells from newly born SD rats massively and purely. Methods SD rats that had been born within 4 to 6 days were used. Their bilateral sciatic nerves were dissected. The nerve fascicles were then extracted under 16× microscope. Two enzymes were used to digest the sciatic nerve specimens twice. G 418 was added in the culture medium for 4 days after 4 days. Results An average number of 3.62× 106 cells were obtained from the sciatic nerves. S 100 stain showed that more than 96% of cultured cells were positive. Conclusion This method can obtain massive purified normal Schwann cells to satisfy the need of tissue engineered bioartificial nerve graft. [