摘要
目的拟建立人胚胎嗅球嗅鞘细胞的培养方法并探讨这些细胞在活性、纯度方面是否具备临床应用水平。方法用含有促进细胞生长的谷氨酰胺、转铁蛋白、生物素、硒酸纳的无血清纯化液纯化嗅鞘细胞,胰蛋白酶消化收集细胞,制成单细胞悬液。采用台盼蓝染色法进行活性测定,P75NGFR和S-100双标免疫荧光染色鉴定,以Hoechst复染鉴定OECs的纯度。结果可见比较典型的嗅鞘细胞:双极或梭形、多突起形和扁圆形,主要以梭形和多突起形细胞为主。细胞活性大于95%,纯度大于90%。结论实验建立的人胚胎嗅球嗅鞘细胞原代培养的方法可行,细胞活性大于95%,纯度和均一性已达到临床应用水平。
Objective To establish a cultural method for human embryonic olfactory ensheathing cells（OECs）of olfactory bulb in order to obtain cells with better viability and purification to satisfy the clinical application.Methods The olfactory ensheathing cells were purified with serum-free purification liquid containing glutamine,transferrin,biotin,sodium selenate,and were digested with trypsinase and collected as the single cell suspension.The cell viability was determined with trypan blue staining,the purity of olfactory ensheathing cells was identified with P75NGFR and S-100 double-labeled immunofluorescence staining.Results More typical olfactory ensheathing cells were found with multi-protruding shape or spindle-shape,less with bipolar or flat circular.The viability of olfactory ensheathing cells was more than 95%,the purity of the cells was more than 90%.Conclusion The cultured olfactory ensheathing cells might be available in clinical research.
出处
《解剖科学进展》
CAS
2010年第2期97-100,共4页
Progress of Anatomical Sciences
基金
国家自然科学基金资助项目(No30971527)
关键词
嗅鞘细胞
细胞培养
纯化
免疫荧光
人胚
olfactory ensheathing cells cell culture purification immunofluorescence human embryo
