为评价PK15细胞能否用于兽用疫苗的研究和生产,对其进行了生物学特性研究和微生物污染及病毒外源因子检测。结果表明:PK15细胞复苏活力为91.77%,呈贴壁性生长,形态为上皮型;生长曲线呈"S"型,最大增殖浓度为24.67×10^4个·mL-1,群体倍增时间为21.08h;同工酶为5条,染色体2n=38,证实为猪源性且没有发生细胞间的交叉污染;细菌、真菌和支原体检查均为阴性;细胞体外培养观察、血吸附试验、接种鸡胚和动物以及特异性荧光抗体结合物检查病毒均为阴性;能较好地表达外源基因,达到了兽用疫苗生产用细胞的质量要求,可为以该细胞为基质的疫苗研究和生产提供细胞资源。
In order to evaluate whether a PK15 cell strain can be used for the research and production of live vac- cine,its biological characteristic,microbial contamination and adventitious virus agents were detected. The resuits showed that the viability of PK15 cells was 91.77% after thawing; the cells were adherent and epithelioid. The growth curve of PK15 cell was S-shape, and the maximum concentration was 24.67 × 10^4 cells-mL^-1 with the population doubling time (PDT) of 21.08 h. Lactic dehydrogenase (LDH) showed 5 bands and karyotyping analysis indicated that the chromosome number of a normal cell was 2n= 38,it demonstrated that it was proeine-derived cell line and had no contamination among cell lines. The cell was free of infections from bacteri- a, fungi and mycoplasmas ; Detection in vitro culture, haemadsorption test, chicken embryo and animal inocula- tion test and determination of special fluorescent combination were all negative. The extro-gene could be effectively expressed. The quality of the cell line conformed to the quality control standands of live vaccine, and it could offer cell resources for the research and production of vaccine based on PK15 ceils.