摘要
目的改良既往新生大鼠视神经组织块培养法体外培养少突胶质细胞. 方法新生2 d SD 大鼠.无菌条件下取双侧视神经.置于预先涂有多聚赖胺酸的培养皿中（直径3..5 cm）.加入基础培养液约400μL.培养第4 天时.更换为含0. 5%胎牛血清化学限定培养液约400 μL.约第10 天时.更换为无胎牛血清化学限定培养液约600 μL.第11 天行髓鞘碱性蛋白（myelin basic protein.MBP）细胞免疫化学鉴定.计算阳性细胞百分率. 重复培养3 次.方差分析方法的稳定性. 结果视神经培养至第11 天.每皿细胞数可达（6-8）×105个.90%以上为MBP 阳性细胞.3 次培养细胞的MBP 免疫细胞化学染色阳性细胞百分率差异无统计学意义（P〉0.05）. 结论本实验方法所得成熟少突胶质细胞纯度高.数量足够-般细胞生物学实验使用.且方法简便、稳定.
Objective To improve the past method for in vitro culture of neonatal rat optic nerve oligodendrocytes. Methods Six optic nerves of newborn rats were placed in poly-lysine precoated dish( diameter 3. 5 cm),cultured with 400 μL basic culture medium for 3 days,and then replaced by 400 μL chemically defined culture medium containing 3% foetal bovine serum( FBS).For about the 10 th days,culture medium was replaced by 600 μL chemically defined culture medium without FBS. At the 11 st days,immunocytochemistry with myelin basic protein( MBP) was used to identify oligodendrocyte,and the percentage of positive cells was calculated. Oligodendrocyte in vitro culture with the method was repeated three times,analysis of variance was used to evaluate the stability of the improved method. Results For the 11 st cultured days,number of cells in per dish was up to( 6 — 8) × 105,more than 90% cells were positive with MBP immunocytochemistry. There was no statistical difference among the percentage of MBP immunocytochemistry positive cells of the three times( P > 0. 05) of culture. Conclusion Higher purified and enough for cell biology experiments mature oligodendrocytes can be obtained with the improved method for in vitro culture of neonatal rat optic nerve oligodendrocytes. This improved method is relatively easy and stable,compared with the other methods for oligodendrocytes culture.
出处
《转化医学杂志》
2014年第6期330-332,346,共4页
Translational Medicine Journal
