改良新生大鼠雪旺细胞的体外培养和纯化

Modified Culture and Purification of Schwann Cells from Neonatal Rats

为建立一种简易有效的体外培养和纯化雪旺细胞(Schwann cells,SCs)的方法,采用显微技术严格无菌条件下取新生大鼠坐骨神经,去除鞘膜,剪碎成0.5 mm3大小,混合酶分步消化法分离单个细胞,用连续阶段时间点(接种培养后第15 min、30 min和45 min)差速贴壁法联合适时阿糖胞苷抑制法进行纯化,通过形态学观察和S-100免疫组化鉴定.结果显示SCs增殖能力强,生长良好,外形多为双极,成堆或旋窝状生长,S-100免疫细胞化学反应阳性细胞纯度均达98%.因此该培养方法可从新生大鼠坐骨神经获得高纯度的雪旺细胞.

To set up a simple and available method of culturing and purifying the Schwann cells（SCs）,the sciatic nerve was obtained from neonatal rats by using microsurgical techniques.Nerve sheath was first removed,afterwards nerve was cut to be 0.5 mm3 tissue pieces,trypsin and collagenase were used to separate SCs by step digestion.Purified SCs was obtained after a differential adhesion at various time points（15 min,30 min and 45 min post-culture） and the treatment of cytarabine（Ara-C） in the course of passage.The purity of SCs was identified through the phase contrast microscope and S-100 immunocytochemical stain.Results show SCs grew in good state as bipolar cells with long and slim enations under inverted phase contrast microscope.Along culturing time prolongation,the SCs grew in heaps or eddy-like.Purity of the SCs with positive immune-cytochemistry of S-100 was exceeded 98% by this culture method.So the culturing method can get high purity Schwann cells from sciatic nerve of neonatal rats.