猪流行性腹泻病毒优势VERO细胞株的克隆和培养OA

[Objective]To enhance the viral titer of porcine epidemic diarrhea virus(PEDV)in culture,this study aimed to obtain a dom-inant VERO cell line through cloning and screening.[Methods]VERO cells were cloned by the limited dilution method over two rounds.Clones exhibiting favorable morphology and rapid growth were preliminarily selected based on microscopic observation.The proliferative capacity of selected clone strains was compared to that of the parental cell line using cell growth curves.The susceptibility of clones to PEDV infection was evaluated using a PEDV infection assay.Optimal culture conditions for highly sensitive clones were determined by conducting experiments with different serum concentrations.[Results]Two VERO cell clones(HB9 and HA3)were ob-tained.Among these,the proliferation rate of the HB9 clone was significantly higher than that of both the parental cells and the HA3 clone.The PEDV titer reached in HB9 cells was as high as 8.00 log10TCID50/mL,significantly exceeding that achieved in the parental cell line or the HA3 clone.Furthermore,the HB9 clone maintained high virus production even at a 6%serum concentration,which was 4%lower than the original culture concentration of 10%.[Conclusion]This study successfully isolated a dominant VERO cell clone(HB9)and optimized its culture conditions,establishing an important foundation for the large-scale production of high-titer PEDV antigen.