成年鼠坐骨神经雪旺细胞体外培养的实验研究

Experimental study od Schwannecell culture from adult SD rats

目的探讨从成年鼠坐骨神经分离培养获得大量雪旺细胞的有效手段。方法取SD大鼠双侧预变性的坐骨神经,预变性后,剪碎至1mm3大小的组织块,单酶消化法进行分离培养,Geneticin液抑制成纤维细胞生长,低浓度酶快速消化传代进一步纯化雪旺细胞,通过相差显微镜活细胞计数和S-100细胞化学标记相结合鉴定了雪旺细胞增值和纯化程度。结果通过上述方法分离培养获得了经S-100蛋白鉴定纯度为85%的第三代雪旺细胞,细胞数量为2.764×107/ml,细胞形态多数为梭形。结论本方法可从大鼠预变性的坐骨神经获得形态与活力良好的雪旺细胞。

Objective To present an effective technique for culture and expansion of Schawann cells from adult SD rats＇ sciatic nerve. Methods SD rats＇ sciatic nerve was pre-degenerated for 10 days. Then SD rats＇ sciatic nerve was dissected. The nerve fascicles were then extracted under 10×microscope. The nerve was cut to be 1mm^3 tissue pieces. One enzyme was used to digest the sciatic nerve speciments. Geneticin was used to be inhibition fibroblasts grow. Low density enzyme quickly digested to passage Schwann cells, the purity of Schawann cells identified through the phase contrast microscope and S-100 immunocytochemical staining. Results Through above-mentioned methods we got high purified Schwann cells.The purity of Schwann cells was 85%.The cell quantity was 2.764× 10^7ml. The morphology of most third passage cells was fusiform. Conclusion Through above-mentioned methods,We can get high quantity Schawann cells,with normal morphology and high vitality.