Gut J, Petersen C, Nelson R, Leech J
Department of Medicine, University of California, San Francisco, San Francisco General Hospital 94110.
J Protozool. 1991 Nov-Dec;38(6):72S-73S.
To facilitate studies of the biology of Cryptosporidium parvum, we have developed an in vitro culture system using Madin-Darby canine kidney (MDCK) cells as the host cell. Oocysts or free sporozoites were incubated 37 degrees C with monolayers of MDCK cells in supplemented RPMI 1640 medium and the cells were examined at various time intervals after initiation of the culture. High rates of infection (up to 90% of MDCK cells) were achievable. Sequential development of trophozoites, meronts, microgametocytes, and macrogametocytes was observed over a 72-h period of culture. Between 72 and 96 h we observed formation of oocyst walls, but fully sporulated oocysts were not observed. This culture system provides access to both the asexual and sexual intracellular stages of C. parvum.
为便于对微小隐孢子虫的生物学特性进行研究,我们开发了一种体外培养系统,该系统使用犬肾传代细胞(MDCK)作为宿主细胞。将卵囊或游离子孢子在补充了RPMI 1640培养基的条件下与MDCK细胞单层在37℃孵育,并在培养开始后的不同时间间隔对细胞进行检查。可实现高感染率(高达90%的MDCK细胞)。在72小时的培养期内观察到滋养体、裂殖体、小配子体和大配子体的连续发育。在72至96小时之间,我们观察到卵囊壁的形成,但未观察到完全孢子化的卵囊。该培养系统可用于研究微小隐孢子虫的无性和有性细胞内阶段。