【摘要】 目的:建立体外培养低代次293细胞的方法。方法:细胞复苏,将冻存的低代次293细胞于37℃水浴中快速融解,移入75cm~2细胞培养瓶中,加入10mL低代次293细胞培养基,6～8h细胞贴壁后更换培养基。细胞传代,吸出培养基,用1×柠檬酸盐细胞消化液洗2遍,留取0.5mL37℃消化5～10min,待细胞脱壁后加入3mL培养基,轻轻吹匀以1∶2比率传代培养。细胞冻存,以1×柠檬酸盐细胞消化液消化5～10min,细胞脱壁后加入5mL细胞培养基中和消化液,离心收集细胞,以1mL的细胞冻存液(90%FBS+10%DMSO)重悬、冻存。结果:细胞传代后8h细胞完全贴壁;以腺病毒感染质粒感染293细胞14d后可观察到腺病毒空斑形成。结论:该方法操作简便,对细胞损伤小且保持细胞生物学特性不变。更多还原

【Abstract】 Objective To establish the cultivated method of low passage 293 ce lls in vivo.Methods Low passage 293 cells were cultivated in vivo according to our methods,at the same time adherence ratio,growth curve and bionomics of low p assage 293 cells were detected.Results 8 hours postpassage,low passage 293 cell s become adherence completely.After fourteen days of the cells posttransfected by the recombant adenovirus,adenovirus plaque was formed in the monolayer of 293 cells.Conclusion The methods established by ourself can maintain vigor and bio nomics of low passage 293 cells.更多还原