非洲绿猴肾细胞(Vero)无血清细胞库的建立

Establishment of Serum-free Vero Cell Banks

目的建立Vero细胞无血清细胞库。方法采用序贯适应和直接适应方法,将Vero细胞从有血清培养适应到无血清培养,建立Vero细胞无血清细胞主库和工作库,采用STR图谱并结合染色体计数对Vero细胞进行鉴别,并对Vero细胞无血清细胞库进行致瘤性和病毒外源因子检定。结果两种方法均能使Vero细胞适应到无血清培养,其中直接适应法可大大缩短适应代次,因此选择其作为建立Vero细胞无血清细胞库的方法。Vero细胞无血清主库和工作库的STR图谱与日本细胞库公布的Vero细胞STR图谱完全一致,Vero细胞库无外源因子污染,无致瘤性。结论Vero细胞无血清细胞库可用作生物制品生产用候选细胞基质。

Objective To establish serum-free Vero cell banks. Methods Establish master and working Vero cell banks by continuous and direct adaptations of Vero cells from serum-containing medium to serum-free medium. Identify the Vero cells by STR pattern and chromosome counting. Test the established cell banks for tumorigenicity and exogenous viral agents. Results Vero cells were successfully adapted to serum-free medium by both continuous and direct adaptations. However,the passage of directly adapted Vero cells decreased as compared with that of continuously adapted ones. Both the STR patterns of established master and working Vero cell banks were completely identical to those of Japan Vero cell banks. No exogenous viral agents or tumorigenicity was observed in the established cell banks. Conclusion The established Vero cell banks may be used as a candidate cell substrate for biologics.