ST贴壁细胞悬浮驯化及应用

Establishment and Application of Suspension Cell Lines of ST Cells

为将ST贴壁细胞通过自主驯化,使其能在ST-S细胞无血清培养基中悬浮生长且能稳定传代,在摇瓶中实现高密度生长,并应用于猪伪狂犬病毒(PRV)悬浮培养,经ST-A低血清培养基适应培养,对一株贴壁的ST细胞进行了无血清的全悬浮驯化,并将PRV在悬浮细胞中连续盲传培养。结果显示:ST悬浮细胞能在无血清培养基中传代,培养48 h至第3代后,所能达到的最终细胞密度为3.00×10^(6 )cells/mL以上;PRV连续培养至第5代,毒价可达到109.0 TCID50/mL。结果表明,用专用培养基可使ST贴壁细胞实现悬浮驯化,并可应用于PRV悬浮培养。本研究为获得高密度ST悬浮细胞和提高PRV增殖效率奠定了技术基础。

In order to make ST adherent cells domesticated by themselves to achieve suspension culture in ST-S serum-free medium as well as stable passage,achieve the growth at high density in shake fl ask,and to use the suspension cell lines for culturing porcine pseudorabies virus(PRV),one strain of ST adherent cell was acclimated within serum-free suspension through adaptive culture in ST-A low serum medium,and PRV was continuously and blindly cultured and passaged within the suspension cells.The results showed that the ST cells could be passaged within serum-free medium,and reach up to the fi nal density of over 3.00×10^(6) cells/mL after 48 hours and to their third generation;the titer of PRV could be up to 109.0 TCID50/mL when it was continuously cultured to its fi fth generation.It was concluded that ST cells could be acclimated under suspension within special medium,and could be used for suspension culture of PRV.A technical foundation was thus laid for obtaining ST suspension cells with high density and for improving PRV reproduction.