新生和成年大鼠雪旺细胞培养、纯化的实验研究

关键词：雪旺细胞 大鼠 坐骨神经 细胞培养 增殖

Keywords:Schwann cells  rat  sciatic nerve  cell culture  proliferation

摘要：

      目的：比较新生和成年大鼠雪旺细胞（Schwann cells，SCs）的体外培养和纯化方法。方法：分别取新生和成年SD大鼠的坐骨神经，用混合酶消化法、差速贴壁法和低浓度胰酶快速消化法进行培养和纯化，通过形态学观察和S-100免疫细胞化学鉴定，并行MTF法检测细胞增殖能力。结果：两组细胞生长状态良好，均呈S-100免疫细胞化学反应阳性，纯度均达95％以上；由生长曲线显示新生鼠SCs生长更快，第2代SCs倍增时间：新生鼠3d、成年鼠4d。结论：从新生和成年大鼠坐骨神经能成功分离培养出大量高纯度的SCs，新生鼠SCs增殖能力更强。

Abstract:

      Objective:To compare the methods of culturing and purifying newborn and adult rat SCs. Method: The sciatic nerves of newborn(aged 1-2 days) and adult(aged 3-4 months) SD rat were harvested. Trypsin and collagenase were used to separate SCs. Purifying SCs was obtained after a differential adhesion and low concentration trypsin in the course of passage. The purity of SCs was identified through the phase contrast microscope and S-100 immunocytochemical stain. The proliferating function of these SCs was assessed by MTT assay. Result: The cells in the two groups were in a good state and S-100-postive by immunocytochemistry, and the purity of SCs was more than 95%. The newborn SCs grew faster than the adult SCs. The doubling time of the 2nd passage of cells was 3 days for the newborn SCs and 4 days for the adult SCs. Conclusion:These findings indicate that a great number of highly purified SCs can be acquired from newborn and adult rat. The proliferation capacity of SCs isolated from newborn rat is higher than those from adult rat.