低血清培养PK-15细胞系对TGEV增殖规律的研究

为降低疫苗生产综合成本,本研究探讨了以低血清培养基(M08011)替代常规细胞培养基(DMEM)的可行性。本研究依次采用含100、80、50、30、20、10、0mL/L血清的低血清培养基(M08011)驯化PK-15细胞,并考察猪传染性胃肠炎病毒(TGEV)在低血清培养的PK-15细胞系中的增殖规律。结果显示,驯化后的PK-15细胞在含100、80、50、30、20 mL/L的低血清培养基中生长状态良好,与常规培养基所培养的PK-15细胞无差异。低血清培养体系培养PK-15细胞时,血清最低添加量为20 mL/L。用该体系培养的PK-15细胞接种TGEV后,TGEV的TCID50为10^-4.97/100μL,与常规条件培养的TGEV的TCID50为104.88/100μL相比,差别不明显。TGEV接种PK-15细胞,低血清培养基中血清浓度降低至20 mL/L,毒价未受到影响。结果表明,本研究建立的PK-15细胞及TGEV低血清培养体系可初步应用于TGEV的增殖培养,为相关疫苗研发奠定了基础。

Study on the proliferation of TGEV by low serum cultured PK-15 cell line

In order to reduce the overall cost of vaccine production in low serum medium(M08011),the feasibility to replace the conventional containing 100μL/L serum concentrations of DMEM medium was observed.The results showed that the growth status of PK-15 cell lines in the low serum medium containing 100,80,50,30 and 20μL/L respectively was not different from that of PK-15 cells cultured in the conventional medium.Therefore,the lowest serum concentration of PK-15 cells was 20μL/L when cultured with the low serum culture system.After inoculated the PK-15 cells with this system for TGEV,the TGEV(TCID50)in the low serum culture was 104.97/100μL,and the difference was not obvious between the normal conditions and the TGEV(TCID50)as 10^-4.88/100μL.The serum concentration of PK-15 cells inoculated with TGEV was reduced to 20 mL/L in low serum medium,and the toxicity was not affected.Therefore,the PK-15 cells and TGEV low serum culture system established in this study can be initially applied to the proliferation and culture of TGEV,laying the foundation for related vaccine research.