成年SD大鼠雪旺细胞在ZQ生物膜上接种及体外培养的研究

Culture of rat Schwann cells derived from degenerated peripheral nerve on ZQ membrane

目的 :探索成年大鼠雪旺 ( SC)细胞的体外培养及与 ZQ膜支架的复合特性。方法 :成年 SD大鼠的坐骨神经作损伤预变性处理 14 d,采用复合酶消化法培养 ,应用差速贴壁、差速消化、及阿糖胞苷孵育 ,进行纯化 ;MTT法计算第二代 SC细胞的生长曲线和倍增时间 ;将 SC细胞接种于 ZQ膜 ,观察细胞在支架表面贴附、生长情况 ;抗 S10 0免疫组织化学方法鉴定 SC细胞。结果 :成年大鼠的坐骨神经作损伤预变性处理 ,采用复合酶消化法培养及纯化可培养出大量具良好增殖活性的 SC细胞 ,纯度可达 95 %以上 ;第二代细胞倍增时间为 3 d;SC细胞在 ZQ膜上能很好贴附生长。结论 :ZQ膜与高纯度可传代的 SC细胞有良好亲和性 ,是一种有应用潜力的外周神经组织工程支架材料。

?Objective: To study the methodology of the culture of Schwann cells derived from degenerated peripheral nerve. Method: Sciatic nerve of adult rats was surgically cut. 14 days after operation, the degenerated nerve tissue was obtained and treated with trypsin and collagenase typeⅡ to prepare single cell suspension,the cells were purified by different speed of attachment and digestion, and incubated on ZQ membrane in the presence of 10 -5 mol/L cytosine arabinoside. The growth of the cells of passage 2 was studied by MTT assay. Schwann cells were identified with anti S100 immumohistochemistry. Results: The cultured cells were spindly in shape and 95% of them were S100 positive. The population doubling time of passage 2 cells was 72 h.The cells attached and stretched on ZQ membrane as well as on the culture vessel surface. Conclusion: Schwann cells can be cultured and purified by predegeneration of the peripheral nerve,different speed of attachment and digestion and the presence of cytosine arabinoside. The cells can grow well on ZQ membrane.