微小隐孢子虫在HCT-8细胞内的培养

目的将人回盲肠癌(HCT-8)细胞作为微小隐孢子虫IId亚型体外感染对象,观察虫体在细胞中的生长发育过程。方法将纯化的隐孢子虫IId亚型感染HCT-8细胞,通过Giemsa染色法观察微小隐孢子虫IId亚型在HCT-8细胞中发育过程,运用PCR方法对不同感染时间点的细胞样品DNA进行检测。结果在感染后72 h内,隐孢子虫出现连续发育阶段,包括脱囊、子孢子、滋养体、裂殖体、配子体、合子和卵囊;PCR的检测结果显示,在感染细胞96 h仍能检测到虫体DNA。结论通过HCT-8细胞,观察到微小隐孢子虫IId亚型的完整的生长发育过程,为抗隐孢子虫药物的筛选提供理论基础,亦可成为微小隐孢子虫IId亚型卵囊体外扩增的方法。

In vitro culture of Cryptosporidium parvum in HCT-8

In order to develop in vitro infected model for Cryptosporidium parvum and to observe its developmental stages in the cells, the human ileocecal cancer (HCT-8) cells were selected to culture the parasite. The purified oocysts of C. parvum IId subtype infected HCT-8 cells in vitro, the developmental processes of C. parvum IId subtype in HCT-8 cells were observed using Giemsa staining, and the DNA of C. parvum IId subtype from different time points of infection were detected by Polymerase Chain Reaction (PCR). Following 72 h co-culture, excystation, sporozoites, trophozoites, meronts, microgametocytes, macrogametocytes, zygote and oocyst appeared orderly. Furthermore, the DNA of C. parvum IId subtype in cells were still detected at 96 h post-infection. The complete developmental processes of C. parvum IId subtype can be observed in HCT-8 cells, which provides a theoretical basis for the screening of anti-Cryptosporidium drugs and becomes a method for C. parvum IId oocyst amplification by in vitro culture.