激活态雪旺细胞在胶元几丁糖膜上生长规律的实验研究

An Experimental Research of the Growth Rule of Activated Schwann cells Cultured on Chitosancollagen film.

目的探索成年SD大鼠激活态雪旺细胞(SC)在体外在胶元几丁糖膜上的生长规律.方法成年SD大鼠的坐骨神经切断预变性7天,采用复合酶分步消化法,接种于胶元几丁糖膜上,通过相差显微镜和扫描电镜观察细胞生长情况,S-100染色鉴定细胞的纯化程度.结果200μl浓度为2×104/ml的激活态的SCs接种于胶元几丁糖膜上和培养皿上,2周后细胞浓度分别达到30×104/ml和20×104/ml,相差显微镜和扫描电镜下观察显示SCs在胶元几丁糖膜上贴壁生长情况良好,体外倍增时间为4天.绘制其生长曲线.结论胶元几丁糖膜和高度纯化的激活态雪旺细胞有良好的亲和性,以其为工艺材料制成的组织工程支架很有可能在促进周围神经再生中发挥优势作用.

ObjectiveTo explore the growth rule of activated SC cultured on the surface of Chitosan- collagen film, derived from adult SD rat. Methods Adult SD rat sciatic nerves were cut and predegenerated for 7 days and SC_s were harvested in a two-step enzymolysis way and cultured on the surface of Chitosan-collagen film. Their growth complexions were observed through the phase contrast microscope and scanning electron microscope. The purity of SC was identified through S-100 munocyto chemical staining. Results 200μl activated SC_s (concentration 2×10~4/ml) were cultured on the surface of Chitosan-collagen film and cell culture dish, the final concentration were up to 30× 10~4/ml and 20×10~4/ml separately after 2 weeks. Under phase contrast microscope and scanning electron microscope, activated SC_s grow well on the surface of the Chitosan-collagen film. The doubling period of SC_s was 4 days. The growth curve was drawled. Conclusion High purified activated SC_s can be cultured on the surface of Chitosan-collagen film. The tissue engineering scaffolds made of it will enhance the regeneration of the peripheral nerve.