东方田鼠皮肤成纤维永生化细胞的建立  

Establishment of an immortalized Microtus fortis skin fibroblast cell line

作　　者：成钢[1] 王京仁[1] 曾文虎[1] 李淑红[1] 王文彬[1] 

机构地区：[1]湖南文理学院动物学湖南省高校重点实验室/环洞庭湖生物资源保育与利用研究中心,湖南常德415000

出　　处：《中国预防兽医学报》2013年第6期453-455,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金：湖南省重点学科建设项目;湖南文理学院省级平台开放项目(SK Y PT201201);博士科研启动项目资助(107/13101046)及科学研究一般项目(300/10301013)

摘　　要：为建立东方田鼠皮肤成纤维细胞(MfSF)系,本研究采用脂质体基因转染法将含有SV40T基因的pSV3neo重组质粒导入原代MfSF中,通过G418筛选,阳性克隆扩大培养,建立永生化M fSF系。实验结果表明,阳性细胞克隆在G418筛选第三周后出现,经扩大培养已稳定传代达42代,而且生长状态良好,PCR及RT-PCR检测结果表明,SV40 T基因已整合到M fSF中并稳定表达。本研究利用SV40T基因导入制备了永生化M fSF细胞系,为动物间成纤维细胞比较研究及相关病原的研究提供了敏感细胞系。In order to establish an immortalized Microtus fortis skin fibroblast （MfSF） line, a mammalian expression recombinant plasmid of pSV3neo containing the SV40 large T antigen gene was used to transfect into the primary MfSF cells using LipofectarnineTM 2000. The positive colonies were appeared after 3 weeks screened by G418, and expanded into immortalized cell lines which passed stably for 42 passages. In addition, the integration of the large T antigen gene into the genome of the immortalized MfSF and the mRNA of SV40 large T antigen in the expanded cells were detected by PCR and RT-PCR, respectively, which substantially proved the immortalized MfSF line was established. The established MfSF line provided a basis for comparative study of fibroblasts fi＇om different animals and the etiological studies in vitro.

关 键 词：东方田鼠 皮肤成纤维细胞 永生化 SV40T抗原 

分 类 号：Q25[生物学—细胞生物学]