【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)少突膠質細胞。方法新生2 d SD大鼠,無菌條件下取雙側視神經(jīng),置于預先涂有多聚賴胺酸的培養(yǎng)皿中(直徑3.5 cm),加入基礎培養(yǎng)液約400μL;培養(yǎng)第4天時,更換為含0.5%胎牛血清化學限定培養(yǎng)液約400μL;約第10天時,更換為無胎牛血清化學限定培養(yǎng)液約600μL;第11天行髓鞘堿性蛋白(myelin basic protein,MBP)細胞免疫化學鑒定,計算陽性細胞百分率。重復培養(yǎng)3次,方差分析方法的穩(wěn)定性。結果視神經(jīng)培養(yǎng)至第11天,每皿細胞數(shù)可達(6~8)×105個,90%以上為MBP陽性細胞;3次培養(yǎng)細胞的MBP免疫細胞化學染色陽性細胞百分率差異無統(tǒng)計學意義(P0.05)。結論本實驗方法所得成熟少突膠質細胞純度高,數(shù)量足夠一般細胞生物學實驗使用,且方法簡便、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)科大學附屬大連市中心醫(yī)院神經(jīng)內(nèi)科;大連醫(yī)科大學;大連醫(yī)科大學附屬大連市中心醫(yī)院綜合神經(jīng)外科;大連醫(yī)科大學附屬大連市中心醫(yī)院康復醫(yī)學科;大連市第三人民醫(yī)院神經(jīng)內(nèi)科;
【分類號】：R329.2

【參考文獻】

相關期刊論文 前5條

1 陳麗萍;郭力;李振飛;平曼;李茹;劉佳;謝小華;董梅;;少突膠質祖細胞的體外分離、培養(yǎng)和定向分化[J];廣東醫(yī)學;2012年15期

2 何平,沈馨亞;少突膠質細胞增殖和分化的研究進展[J];神經(jīng)解剖學雜志;2000年02期

3 薛慶善，，郭畹華;大鼠大腦皮質星形膠質細胞的體外培養(yǎng)及其促神經(jīng)突起生長作用研究[J];神經(jīng)解剖學雜志;1996年02期

4 彭錫嘉,劉少章,葉劍,袁容娣;新生大鼠視神經(jīng)少突膠質細胞體外培養(yǎng)純化及鑒定[J];眼科新進展;2003年03期

5 陳曉靜;謝敏杰;;新生SD大鼠少突膠質細胞前體細胞的培養(yǎng)與分化[J];卒中與神經(jīng)疾病;2014年01期

【共引文獻】

相關期刊論文 前10條

1 王秀文;張立霞;楊成;潘晶晶;王紅;;電針對體外培養(yǎng)星形膠質細胞機械性損傷后反應性膠質化的影響[J];濱州醫(yī)學院學報;2010年04期

2 劉蘋,彭錫嘉,葉劍,袁容娣;新生大鼠視神經(jīng)少突膠質細胞原代培養(yǎng)及鑒定[J];重慶醫(yī)學;2003年03期

3 徐錫金,樸英杰,霍霞;人發(fā)角蛋白對脊髓損傷大鼠少突膠質細胞增殖分化效應的影響[J];第一軍醫(yī)大學學報;2003年06期

4 殷紅兵;林航;吳衛(wèi)平;穆軍山;葉建新;林敏;林曉姝;武雷;姚麗青;;間歇性低氧對少突膠質前體細胞增殖的影響[J];福州總醫(yī)院學報;2007年Z1期

5 殷紅兵;吳衛(wèi)平;林航;穆軍山;葉建新;林敏;林曉姝;范明;;細胞因子對少突膠質細胞增殖的影響[J];福州總醫(yī)院學報;2007年Z1期

6 馬麗香,李瑞錫,王R

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