人脑胶质瘤细胞的原代培养及生长活性观察.pdfVIP

( Ch in J Neurosurg D is Res) 2005; 4( 3) # 245# : 167 1- 2897( 2005) 04- 245- 03 # # 易伟 牛洪泉 陶胜忠 叶飞 雷霆 薛德麟 ( , 430030) , 24, + + , ( PCNA) ( MTT ) 20, 4 98% , PCNAMTT , + + ; , ; MTT PCNA ; ; ( PCNA); MTT R 329. 2 A Character ization and growth activity observation of prim ary hum an gliom as cell culture YI We,i N IUH ong quan, TA O Shengzhong, YE Fe,i LEI T ing, X UE D elin D ep ar tm ent o N eurosurg ery, Tongj i H osp ital, Tongj i M edical Colleg e, H uazhong Univ ersity o Sc ience and Technology, W uhan 430030, China Abstract Obj ective T o in estigate the m ethods of prmi ary hum an gliomas cell culture and the relationsh ip bewt een the cu ltured cells and their histological grades. M ethods Twenty-four prmi ary gliom asw ere cultured w ith enzym atic d igestion and the tumor cells were purifiedw ith a new protocol. The processw as d igesting the cells w ith trypsin at first, then oscillating the cells while they adhered, and mechan ically scraping the mi puri- ty cells off if necessary at final. In order to obser e the cells proliferation, methyl thiagolyl tetragolium (MTT) assay and proliferating cell nuclear antigen ( PCNA) mi munoh istochem ical staining were applied. ResultsTwenty prmi ary gliom as were successfully cultured. The new purification protocol could purify the tumour cells beyond 98% . PCNA and MTT assay demonstrated the proliferation of the cu ltured cells correlated w ith their h istological grades. Conclusion Themethod of digestion cu lti ation can successfully culture prmi ary gliomas. Using the puri- fication method of digestion-oscillation-scrape can obtain a h i