大鼠视神经少突胶质细胞培养方法的改良

发布时间：2018-08-11 18:38

【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400μL;约第10天时,更换为无胎牛血清化学限定培养液约600μL;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(P0.05)。结论本实验方法所得成熟少突胶质细胞纯度高,数量足够一般细胞生物学实验使用,且方法简便、稳定。
[Abstract]:Objective to culture oligodendrocytes in vitro by modified method of optic nerve tissue mass culture in neonatal rats. Methods the bilateral optic nerves were taken from SD rats on the second day after birth. The bilateral optic nerves were removed under aseptic condition and placed in a petri dish coated with polylysidic acid (3.5 cm), in diameter and 400 渭 L in the basic culture medium). It was replaced with 0.5% fetal bovine serum chemically limited medium about 400 渭 L on the 10th day, about 600 渭 L on the 10th day, and the myelin basic protein (MBP) was identified by cell immunocytochemistry on the 11th day, and the percentage of positive cells was calculated. The stability of ANOVA was obtained by repeated culture for 3 times. Results there was no significant difference in the percentage of MBP immunocytochemical staining positive cells between the three times of MBP positive cells and the number of MBP immunocytochemistry positive cells in the optic nerve culture on the 11th day (6% 脳 10 5 cells per dish) (P0.05). The results showed that there was no significant difference in the percentage of MBP immunocytochemical staining positive cells between the two groups (P0.05). Conclusion the mature oligodendrocytes obtained by this method are of high purity and sufficient number for general cell biology experiments, and the method is simple and stable.
【作者单位】：大连医科大学附属大连市中心医院神经内科;大连医科大学;大连医科大学附属大连市中心医院综合神经外科;大连医科大学附属大连市中心医院康复医学科;大连市第三人民医院神经内科;
【分类号】：R329.2

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