新孢子虫速殖子在HCT-8、Vero和Hela细胞中培养的比较  

Comparison of Neospora caninumtachyzoites cultured in HCT-8,Vero and Hela cells

作　　者：边征征 吕强[1] 宫鹏涛[1] 常乐[1] 邢沈阳[1] 张西臣[1] 李建华[1] 

机构地区：[1]吉林大学动物医学学院,吉林长春130062

出　　处：《中国病原生物学杂志》2014年第8期721-723,726,共4页Journal of Pathogen Biology

基　　金：国家重点基础研究发展计划项目(2015CB150300);吉林省科技发展计划项目(No.20100222;No.20130521005JH)

摘　　要：目的观察和比较新孢子虫速殖子在人结肠癌细胞(HCT-8)、人宫颈癌细胞(Hela)和非洲绿猿肾细胞(Vero)中的生长情况。方法分别用HCT-8、Hela和Vero细胞(RPMI-1640培养基)连续培养新孢子虫速殖子9d,观察并计数速殖子数量,绘制生长曲线;4d取培养物作吖啶噔染色,用激光共聚焦显微镜观察速殖子。结果新孢子虫速殖子用HCT-8、Vero和Hela细胞培养均生长,尤以在HCT-8和Vero细胞中生长较快,第6d虫体数达到高峰,且在HCT-8中高峰期可持续2d,在Vero中生长高峰可持续1d。在Hela细胞中速殖子生长较慢,虫体数量一直处于较低水平。吖啶噔染色观察,HCT-8和Vero细胞中纳虫泡较大且速殖子数量较多,Hela细胞内纳虫泡较小且速殖子数量较少。结论新孢子虫速殖子在HCT-8细胞中生长较快,纳虫泡较大且速殖子数量较多。HCT-8细胞可以替代Vero细胞用于速殖子的体外培养。Objective To compare the in vitro growth characteristics of tachyzoites of Neospora caninumin the HCT-8human colon cancer cell line,the Hela human cervical carcinoma cell line,and the Vero green monkey kidney cell lineMethods N.caninumtachyzoites were cultured for 9days in RPMI-1640 with HCT-8,Hela,and Vero cells.The growth of these cells was observed and the number of tachyzoites in each set of cells was counted at different times to produce growth curves.Intracellular tachyzoites were also stained with acridine orange on day 4and observed under a laser scanning confocal microscope. Results N.caninumttachyzoites grew in all three cell lines.However,N.caninum grew faster in HCT-8and Vero cells.The number of tachyzoites peaked on day 6.This peak remained in HCT-8cells for2 days and in Vero cells for 1day.N.caninumgrew slower in Hela cells and there were fewer parasites.Results of acridine orange staining indicated that parasitophorous vacuoles in HCT-8and Vero cells were larger than those in Hela.In addition,the number of parasites in HCT-8and Vero cells was greater than that in Hela. Conclusion N.caninum tachyzoites cultured in HCT-8cells grew rapidly with large parasitophorous vacuoles and in large numbers,suggesting that HCT-8could be used as an alternative cell line for the in vitro culture of N.caninumtachyzoites

关 键 词：新孢子虫 速殖子 HCT-8 体外培养 

分 类 号：R382.32[医药卫生—医学寄生虫学]