安氏隐孢子虫在HCT-8和AGS细胞中增殖效果的研究  

In vitro proliferation of Cryptosporidium andersoni in HCT-8 and AGS cell lines

作　　者：逯兆喜[1] 吴亮[1] 姜旭淦[1] 沈玉娟[2] 傅行礼[1] 涂国华[1] 李礼[1] 陈盛霞[1] 曹建平[2] 

机构地区：[1]江苏大学基础医学与医学技术学院,镇江212013 [2]中国疾病预防控制中心寄生虫病预防控制所,卫生部寄生虫病原与媒介生物学重点实验室,世界卫生组织疟疾、血吸虫病和丝虫病合作中心,上海200025

出　　处：《中国人兽共患病学报》2011年第6期475-478,483,共5页Chinese Journal of Zoonoses

基　　金：卫生部寄生虫病原与媒介生物学重点实验室开放课题(No.WS-BKFKT200701);国家卫生行业科研专项(No.200802012);国家传染病重大科技专项(No.2008ZX10004-002,No.2009ZX10004-201);江苏大学科技创新团队项目(No.2008-018-02)联合资助

摘　　要：目的比较安氏隐孢子虫(Cryptosporidium andersoni)在人结肠腺癌(human ileocecal adenocarcinoma,HCT-8)细胞和人胃腺癌(human stomach adenocarcinoma,AGS)细胞中的增殖。方法取培养至对数生长期的HCT-8细胞和AGS细胞,接种于6孔培养板中,待细胞生长融合至60%～70%时,加入2.5×105个安氏隐孢子虫卵囊,培养至24 h和48 h,以Giemsa染色法和Real-time PCR技术检测虫体在细胞中的增殖。结果 Giemsa染色法观察24 h和48 h HCT-8细胞中虫体数量均高于AGS细胞(P<0.05)。Real-time PCR技术测得24 h和48 h HCT-8细胞中安氏隐孢子虫卵囊COWP基因拷贝数均高于AGS细胞(P<0.05)。结论与AGS细胞相比,以HCT-8细胞作为体外感染模型更利于安氏隐孢子虫的增殖。In order to study the proliferation of Cryptosporidium andersoni in human ileocecal adenocarcinoma （HCT-8） cells and human stomach adenoearcinoma （AGS） ceils, HCT-8 and AGS cells were harvested at mid-exponential phase and planted in 6 well plates. When the monolayers reached 60 %- 70 % confluency, the excysted oocysts （2.5× 10^5/well） were inoculated into cells and parasite growth was terminated at 24 h and 48 h. Then the proliferation of parasites was determined by Giemsa staining and Real-time polymerase chain reaction （PCR） assay. At 24 h and 48 h, the number of parasites in HCT-8 cells was higher than that of AGS cells （P^0.05） by Giemsa staining. The copies of COWP gene in HCT-8 cells were also higher than those of AGS cells （P〈0.05） by Real-time PCR assay. Ifs found that the HCT-8 cell line was a more suitable host model to support Cryptosporidium andersoni proliferation than that of AGS cell line.

关 键 词：安氏隐孢子虫 HCT-8细胞 AGS细胞 GIEMSA染色 Real—time PCR 

分 类 号：R382.3[医药卫生—医学寄生虫学]