【摘要】：背景:神經(jīng)元是有絲分裂后的細(xì)胞,體外培養(yǎng)很難存活。脊髓運動神經(jīng)元的分離、純化和培養(yǎng)同時也是細(xì)胞培養(yǎng)的技術(shù)難點。目的:建立新生大鼠脊髓運動神經(jīng)元培養(yǎng)體系,并予以分類鑒定和測定其純度。方法:取新生大鼠脊髓腹側(cè)組織分離成細(xì)胞懸液,經(jīng)密度梯度離心及差速貼壁后純化培養(yǎng),采用免疫細(xì)胞化學(xué)雙標(biāo)染色法對培養(yǎng)蓋片上的細(xì)胞予以鑒定、分類,結(jié)合Hoechst熒光染核,計數(shù)各細(xì)胞成分的含量。結(jié)果與結(jié)論:細(xì)胞貼壁生長良好,神經(jīng)元占85.8%,其中運動神經(jīng)元達(dá)71.6%,星形膠質(zhì)細(xì)胞占7.8%,NF200和膠質(zhì)纖維酸性蛋白染色皆為陰性的細(xì)胞占6.4%。結(jié)果說明,取新生大鼠腹側(cè)脊髓組織結(jié)合密度梯度離心及差速貼壁接種法可以培養(yǎng)出高純度的脊髓運動神經(jīng)元。
[Abstract]:Background: neurons are mitotic cells that are difficult to survive in vitro. The isolation, purification and culture of spinal motoneurons are also the technical difficulties in the culture of spinal cord motoneurons. Objective: to establish the culture system of spinal motoneurons in neonatal rats and to classify and determine its purity. Methods: cell suspension was isolated from ventral tissue of spinal cord of newborn rats and purified and cultured by density gradient centrifugation and differential adhesion. The cells on the culture cover were identified and classified by immunocytochemical double labeling method. Combined with Hoechst fluorescence staining, the contents of cell components were counted. Results & conclusion: the adherent cells grew well, with 85.8% of neurons, 71.6% of motoneurons, 7.8% of astrocytes, 6.4% of cells negative for NF200 and glial fibrillary acidic protein (GFAP) staining, and 71.6% for motor neurons, 7.8% for astrocytes and 6.4% for NF200 and GFAP negative cells. The results showed that the spinal cord motoneurons of high purity could be cultured by density gradient centrifugation and differential adherent inoculation in ventral spinal cord of newborn rats.
【作者單位】： 大連醫(yī)科大學(xué)/大連醫(yī)科大學(xué)附屬第一醫(yī)院;
【基金】：遼寧省自然科學(xué)基金(2013023035) 大連市科技計劃項目(2013E15SF171)~~
【分類號】：R329.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 孫毅;趙冬梅;張燁;劉洪付;黃飛;;胎鼠前角運動神經(jīng)元的取材與培養(yǎng)研究[J];濱州醫(yī)學(xué)院學(xué)報;2009年06期

2 楊曉玲;趙合慶;嚴(yán)志強(qiáng);;體外培養(yǎng)大鼠脊髓運動神經(jīng)元的存活率及形態(tài)觀察[J];基礎(chǔ)醫(yī)學(xué)與臨床;2007年03期

3 陳波;袁瓊蘭;;膠質(zhì)細(xì)胞源性神經(jīng)營養(yǎng)因子與突觸可塑性[J];四川解剖學(xué)雜志;2006年04期

4 趙合慶,楊曉玲,嚴(yán)志強(qiáng);胚胎大鼠脊髓運動神經(jīng)元的體外培養(yǎng)與鑒定[J];中華神經(jīng)科雜志;2004年05期

5 宋學(xué)琴,王麗琴,吳淑玉,李春巖;生長底物對培養(yǎng)胚胎大鼠脊髓運動神經(jīng)元的作用[J];中國免疫學(xué)雜志;2004年06期

6 宋學(xué)琴,吳淑玉,王麗琴,王曉絹,李春巖;胚胎大鼠脊髓運動神經(jīng)元體外培養(yǎng)方法的優(yōu)化[J];河北醫(yī)科大學(xué)學(xué)報;2004年03期

7 宋學(xué)琴,李春巖,吳淑玉,王麗琴,王曉娟;胚胎大鼠脊髓運動神經(jīng)元的體外培養(yǎng)及鑒定[J];基礎(chǔ)醫(yī)學(xué)與臨床;2004年01期

8 余磊,潘樹義,秦建強(qiáng);脊髓神經(jīng)細(xì)胞培養(yǎng)的方法[J];中國臨床解剖學(xué)雜志;2002年04期

9 楊浩,春婷,鞠躬;不同方法純化體外培養(yǎng)脊髓神經(jīng)元的比較[J];中國組織化學(xué)與細(xì)胞化學(xué)雜志;2000年03期

10 楊浩,梁U，

本文編號：2436809