成年大鼠雪旺细胞的快速扩增

EXPANSION OF ADULT SCHWANN CELLS FROM RAT PERIPHERAL NERVES

采用接种雪旺细胞的可降解导管修复外周神经损伤是一种有望替代自体神经移植的方法。如何在短期内利用病人少量的神经碎片获得大量雪旺细胞是该方法用于临床的关键。以大鼠坐骨神经为模型,利用雪旺细胞增殖的内在机制,探索出一种快速增殖成年雪旺细胞的方法。采用预变性7 d的坐骨神经,用酶消化分离出雪旺细胞,接种在层粘连蛋白包被的培养瓶中,经过7 d的培养,获得纯度为96%、细胞密度为600 个/mm2的雪旺细胞,雪旺细胞的纯度和密度明显高于对照的新鲜神经。未使用霍乱毒素、毛喉素等促有丝分裂剂和抑制成纤维细胞的基因毒素,符合临床使用要求。结果表明, 可以利用少量的损伤神经碎片在短期内获得大量可用于临床的雪旺细胞。

Using an artificial nerve conduit containing Schwann cells to repair peripheral nerve injuries is one of the most promising methods, in attempts to design an alternative to autografts. It is necessary for the clinical use of the method to obtain a large number of viable Schwann cells from small amounts of adult nerves fragments within a short period of time. An effective technique for isolation and expansion of Schwann cells from adult rat peripheral nerves was presented. Schwann cells were isolated from the 7 d predegenerated sciatic nerve and seeded on the flasks coated with laminin. After the 7 d proliferation exploiting endogenous mechanisms of cell expansion, the purity of Schwann cells was 96% and the density was about 600 cells/mm2, which were markedly higher than that of Schwann cells isolated from the fresh nerve. Mitogenes and cytotoxic drugs were not used in this method, which meets the demand of clinical application. These results indicated that this method can be used to isolate and expand adult Schwann cells from small amounts of preinjured nerve fragments in a short time period.