Korean Journal of Veterinary Research 1995;35(2):317-326.
Development of Cryptosporidium parvum in cell culture
Bo-sook Kim1, Hoo-don Joo2, Sung-hwan Wee2, Tae-jong Kim3
1Seoul Grand Park
2Department of Parasitology, Veterinary Research Institute of RDA
3Department of Veterinary Medicine, Kon-Kuk University
세포배양에서 Cryptosporidium parvum의 발육
김보숙1, 주후돈2, 위성환2, 김태종3
1서울대공원
2가축위생연구소 기생충학
3건국대학교
Abstract
The purpose of this study was to establish a method for in vitro culture of C parvum isolated in Korea by determination of suitable cell model to complete development of this parasite. The result obtained were summerized as follows: 1. To determine the most suitable cell line, six types of cell line were examined by microscopy. All cell lines were infected with C parvum and showed the highest infection score in HmLu cells. 2. The staining methods including DMSO-modified acid-fast(A-F) stain, hematoxylin-eosin(H & E) stain and immunofluorescence antibody(IFA) stain were applied to examine the infection of C parvum in cell culture. These staining methods were possible to examine the infection of C parvum in cell culture. The most sensitive one was IFA staining technique. 3. Developmental stages of C parvum in HmLu cell were observed. After the initial 8 hour incubation period, some trophozoites were observed. The meronts and gametes were appeared at 24-48 hour post inoculation(PI), and oocysts were observed firstly at 48-72 hour PI. 4. In H & E stain, the parasite appeared as basophilic within parasitophorous vacuole membrane(PVM) and lying in cytoplasm at near the nucleus of the host cells. It was able to distinguish the type I, type II meronts and gametes. 5. In DMSO-modified acid-fast stain, specific stained parasites were appeared firstly after 48 hour PI. The parasites were showed with different degrees of staining bright red color within PVM. 6. The endogenous stages of parasites in HmLu cell recovered at 48, 96, 120 and 144 hour after inoculation were reacted with rabbit immunized serum in immunofluorescence antibody and avidin-biotin complex peroxidase staining technique.
Key Words: C parvum, cell culture, development stages


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