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- 2017
Hi5-SF悬浮细胞系的建立及鉴定
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Abstract:
从中国典型培养物保藏中心(CCTCC)引进High Five细胞并对其进行驯化,获得无血清悬浮驯化株(Hi5-SF)。依据《中华人民共和国药典(2010年版三部)》对其进行细胞生物学特性研究、微生物污染、病毒外源因子及细胞致瘤性检测。结果发现,Hi5-SF细胞复苏活力76.3%,生长曲线呈“S”型,最大增殖密度可达3.50×106 mL-1,群体倍增时间为26.2 h;细菌、真菌及支原体检查均为阴性;血细胞吸附试验、致细胞病变试验和特异性病毒荧光抗体结合物检查结果均为阴性;致瘤性检查结果也为阴性。说明该细胞株符合兽用疫苗生产用细胞质量要求,可以在疫苗研究与生产中应用。
High Five cells were introduced from CCTCC and domestecated,and we obtained semi-adherent cells to suspension growth cell line (Hi5-SF).Subsequently,the biological characteristics,microbial contamination,viral exogenous factors and tumorigenicity of Hi5-SF cell line were detected according to the Pharmacopoeia of People’s Republic of China (3rd Edition,2010).The results showed that the recovery vitality of Hi5-SF cells was 76.3%,the growth curve was S-shape,and maximum concentration was 3.50×106 cells·mL-1 with the population doubling time(PDT) of 26.2 h. The cells were free of bacteria,fungi and mycoplasmas;hemadsorption test,cytopathic test,specific virus fluorescent antibody binding test and tumorigenicity test were all negative. The resultus indicated that the Hi5-SF cell line meets the quality requirements of veterinary vaccine production.