【摘要】：目的改良既往新生大鼠視神經(jīng)組織塊培養(yǎng)法體外培養(yǎng)少突膠質(zhì)細(xì)胞。方法新生2 d SD大鼠,無(wú)菌條件下取雙側(cè)視神經(jīng),置于預(yù)先涂有多聚賴胺酸的培養(yǎng)皿中(直徑3.5 cm),加入基礎(chǔ)培養(yǎng)液約400μL;培養(yǎng)第4天時(shí),更換為含0.5%胎牛血清化學(xué)限定培養(yǎng)液約400μL;約第10天時(shí),更換為無(wú)胎牛血清化學(xué)限定培養(yǎng)液約600μL;第11天行髓鞘堿性蛋白(myelin basic protein,MBP)細(xì)胞免疫化學(xué)鑒定,計(jì)算陽(yáng)性細(xì)胞百分率。重復(fù)培養(yǎng)3次,方差分析方法的穩(wěn)定性。結(jié)果視神經(jīng)培養(yǎng)至第11天,每皿細(xì)胞數(shù)可達(dá)(6~8)×105個(gè),90%以上為MBP陽(yáng)性細(xì)胞;3次培養(yǎng)細(xì)胞的MBP免疫細(xì)胞化學(xué)染色陽(yáng)性細(xì)胞百分率差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論本實(shí)驗(yàn)方法所得成熟少突膠質(zhì)細(xì)胞純度高,數(shù)量足夠一般細(xì)胞生物學(xué)實(shí)驗(yàn)使用,且方法簡(jiǎn)便、穩(wěn)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.
【作者單位】： 大連醫(yī)科大學(xué)附屬大連市中心醫(yī)院神經(jīng)內(nèi)科;大連醫(yī)科大學(xué);大連醫(yī)科大學(xué)附屬大連市中心醫(yī)院綜合神經(jīng)外科;大連醫(yī)科大學(xué)附屬大連市中心醫(yī)院康復(fù)醫(yī)學(xué)科;大連市第三人民醫(yī)院神經(jīng)內(nèi)科;
【分類號(hào)】：R329.2

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本文編號(hào)：2177874