兔腰椎间盘髓核细胞活力测定及外源性基因在其中的表达  

Cells culture of rabbit lumbar nucleus pulposus and the expression of EGF Pgene in these cells

作　　者：肖剑[1] 贾连顺[1] 程岚[2] 潘欣[3] 陈德玉[1] 潘卫[3] 戚中田[3] 

机构地区：[1]解放军第二军医大学长征医院,上海200003 [2]解放军第二军医大学新华医院,上海200092 [3]解放军第二军医大学微生物教研室,上海200000

出　　处：《中国临床康复》2002年第20期3023-3024,T001,共3页Chinese Journal of Clinical Rehabilitation

摘　　要：目的通过对兔腰椎间盘髓核细胞的原代培养,了解髓核细胞的生物学特性,并将绿色荧光蛋白基因转染至髓核内,观察基因在细胞内的表达情况。方法将兔腰椎间盘髓核组织剪碎,并分离出细胞,放入含10%灭活胎牛血清的F12-DMEM培养液中进行培养,建立体外髓核细胞培养模型,同时进行细胞活力测定,并用脂质体转染法将绿色荧光蛋白基因转染至髓核细胞内,观察基因的表达情况。结果(1)兔髓核细胞贴壁及生长的速度均十分缓慢。(2)活力测定提示随着培养时间的延长及传代,活力逐渐降低。(3)转染了绿色荧光蛋白基因的髓核细胞可以持续发出绿色荧光达3个月。结论(1)兔腰椎间盘髓核细胞的生长能力较弱。(2)外源性的基因可以在髓核细胞内得到持续的表达。Objective Rabbit nucleus pulposus cells were c ultured in monolayer,the biologica l action of the cells were observed,a nd the enhanced green fluorescent protein(EGFP)gene was transduced into the cells,a nd the expression of the exogenous ma rker gene was detect-ed.Methods Lumbar nucleus pulposus cells were c ultured in F12-DMEM medium containi ng 10%fetal bovine serum to set up a ce lls cul-turing model,the cells viability wa s assessed.The EGFP gene was transduced into the cells by Lipofectamine,and the cells were observed to see whether them could give out green light.Results The amount of the cells cultured in vitro increased slowly.With the cells culturing extended,the viability of the cells declined.After the EGFP gene was tra nsduced into the cells,the green lig ht could be observed the next day,and the cells could give out light for three month until they dead.Conclusion The growing ability of the nucleus pu lposus cells was feeble.The successful expression of EGFP gene d emonstrated that the exogenous gene can be expressed in nucleus pulposus cell.

关 键 词：腰椎间盘 髓核细胞 活力 测定 外源性基因 绿色荧光蛋白 基因表达 

分 类 号：R681.5[医药卫生—骨科学]