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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。
Objective To compare the proliferation of Cryptosporidium andersoni in human ileocecal adenocarcinoma (HCT-8) cells and human stomach adenocarcinoma (AGS) cells. Methods HCT-8 cells and AGS cells cultured in logarithmic growth phase were inoculated into 6-well culture plates. When the cells grew to 60% -70% confluence, 2.5 × 10 5 Cryptosporidium parvum oocysts were added, After 24 h and 48 h cultivation, the proliferation of the parasite in the cells was detected by Giemsa staining and Real-time PCR. Results The Giemsa staining showed that the numbers of parasites in HCT-8 cells at 24h and 48h were higher than that of AGS cells (P <0.05). The copy number of COWP gene in Cryptosporidium parvum oocysts in HCT-8 cells was higher than that in AGS cells (P <0.05) by Real-time PCR. Conclusion Compared with AGS cells, HCT-8 cells as an in vitro infection model is more conducive to Cryptosporidium parvum proliferation.