摘要
目的:探讨新生大鼠嗅鞘细胞的分离、培养和纯化方法。方法:取2 d内的新生大鼠嗅球组织,在解剖显微镜下分离取材,原代培养嗅鞘细胞,运用差速贴壁法和差速贴壁+阿糖胞苷+细胞生长刺激因子法两种方法进行纯化,在倒置显微镜下观察不同培养时间嗅鞘细胞的形态、数量和NGFRp75抗体免疫荧光结果,统计嗅鞘细胞的纯度。结果:嗅鞘细胞的纯度分别是差速贴壁组平均为60.41%±4.32%,差速贴壁+阿糖胞苷+细胞生长刺激因子法组平均为84.98%±4.03%。两组数据用t检验进行统计学分析,差异有统计学意义(P<0.05)。结论:取材于新生大鼠;显微镜下分离脑膜;差速贴壁+阿糖胞苷+细胞生长刺激因子法纯化,都是良好的嗅鞘细胞培养方法。
Objective：To explore the methods of separation,cultivation and purification of olfactory ensheathing cells（OECs） in neonatal rats.Methods： The olfactory bulb organizations of neonatal rat within 2 days were separated under microscope.The olfactory ensheathing cells were clutured by primary culture,purified by two methods inclouding selective plating technique and selective plating technique＋cytarabine＋cell growth stimulates factor.The quantity and the form of different cultural time olfactory ensheathing cells were observed with inverted microscope,and NGFRp75 immunofluorescent histochemical were observed with fluorescence microscope.Olfactory ensheathing cells purity of two methods were analysed statistically.Results： The purity of olfactory ensheathing cells were 60.41%±4.32% in group of selective plating technique,and 84.98%±4.03% in group of selective plating technique＋cytarabine＋cell growth stimulates factor average.Two sets of data were analysed by t-test,showing significant difference（P0.05）.Conclusion： Drawing materials from the neonatal rats,separating meningeal with microscope,purifying with selective plating technique＋cytarabine＋cell growth stimulates factor were good olfactory ensheathing cells culture method.
出处
《神经解剖学杂志》
CAS
CSCD
北大核心
2012年第3期307-311,共5页
Chinese Journal of Neuroanatomy
基金
湖北省自然科学基金(2009CDB236)
关键词
新生大鼠
嗅鞘细胞
培养
纯化
分离
neonatal rats
OECs
cultivation
purification
separation
