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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 investigate a simple and reliable method for cell culture of large numbers of pure and viable Schwann cells. Methods SD rats of 4 ~ 6 days old were used to dissect the bilateral sciatic nerve and remove the epineurium by dissecting microscope to obtain the nerve bundle. The cells were cut and digested with double digestion method. The cultured cells were purified by G-418 after cultured in DMEM for 4 days. After 4 days, the cells were counted, viability and immunocytochemistry. Results The method can extract 3.62 × 106 Schwann cells from the bilateral sciatic nerve of neonatal rats. After purification, S-100 protein immunocytochemical staining showed that the purity of Schwann cells can reach more than 96%. Cell vitality, after 8d can be cultured. Conclusion This method can obtain a large number of pure Schwann cells with high purity and vitality and meet the needs of tissue engineering artificial nerves.