摘要
目的比较一次性培养瓶与方瓶培养人二倍体细胞2BS株的效果。方法接种相同浓度的人二倍体细胞2BS株至一次性培养瓶和小方瓶中,(37±0.5)℃静置培养3 d,于显微镜下观察细胞生长状态,并进行细胞计数。用0.025～0.05 MOI水痘-带状疱疹病毒Oka株感染2BS细胞,(35±0.5)℃静置培养3 d,待细胞病变达70%以上时,收获病变细胞,制备冻干水痘减毒活疫苗,并参考水痘减毒活疫苗注册标准及《中国药典》(三部)2010版,进行各项指标检定。结果在接种2BS细胞浓度相同的情况下,与方瓶相比,一次性培养瓶可增加一定的细胞培养面积,细胞总数相对较高;一次性培养瓶培养的2BS细胞贴壁速度较快,呈梭形,密度高,方向性好,轮廓清晰,分布均匀,胞质饱满,而方瓶培养的2BS细胞贴壁速度较慢,生长不均匀;使用一次性培养瓶及方瓶培养2BS细胞生产的水痘减毒活疫苗,各项检定指标结果均符合《中国药典》(三部)2010版相关要求,在病毒滴度无明显差异的情况下,使用一次性培养瓶可使水痘疫苗产量大幅度增加。结论使用一次性培养瓶培养人二倍体细胞2BS株生产的水痘减毒活疫苗疫苗产量较方瓶有较大幅度的提高。
Objective To compare the culture effects of human diploid cell 2BS strain with disposable culture flask and with common flask. Methods The 2BS cells at the same concentrations were inoculated into disposable culture flask and small common flask respectively, and incubated at （37 ±0. 5 ） ℃ for 3 d, observed for growth under microscope and count- ed, then infected with varicella-zoster virus （VZV） Oka strain at a MOI of 0. 025 - 0. 05 and incubated at （35 ± 0. 5 ） ℃ for 3 d. The cells were harvested when CPE reached more than 70% to prepare freeze-dried live attenuated vari- eella vaccine. Overall control tests were performed on the prepared vaccine according to the standard for registration of live attenuated varicella vaccine and the requirements in Chinese Pharmacopoeia（Volume Ⅲ, 2010 edition）. Results At the same concentrations for inoculation, the area of 2BS cell culture in disposable culture flask increased as compared with that in common flask, while the total cell count was relatively large. The 2BS cells in disposable flask became adher- ent rapidly, which were in spindle shape, at high density, with good directivity, clear outline and full cytoplasm, and in even distribution. However, the 2BS cells in common flask became adherent slowly and were uneven in growth. All the quality indexes of vaccines prepared with 2BS cells cultured in both disposable culture flask and common flask met the re- quirements in Chinese Pharmacopoeia （Volume Ⅲ, 2010 edition）. If the virus titers showed no significant difference, the yield of varicella vaccine prepared with 2BS cells cultured in disposable culture flask increased remarkably. Conclusion The yield of live attenuated varieella vaccine prepared with 2BS cells cultured in disposable culture flask increased remarkably as compared with that in common flask.
出处
《中国生物制品学杂志》
CAS
CSCD
2013年第10期1376-1378,共3页
Chinese Journal of Biologicals
关键词
一次性培养瓶
人二倍体细胞
水痘减毒活疫苗
Disposable culture flask
Human diploid cells
Live attenuated varicella vaccine
