Journal article
Simplified methods for obtaining purified oocysts from mice and for growing Cryptosporidium parvum in vitro
The Journal of Parasitology, Vol.82(5), pp.757-762
1996
Abstract
Seven- to 8-day-old Arc/Swiss mice were infected with 100,000-120,000 Cryptosporidium parvum oocysts. At 8 days postinfection (PI) the jejunum, ileum, cecum, colon, and rectum were removed. Using a simple extraction procedure and purification by Ficoll gradient centrifugation, we rountinely obtained between 3-6 million and up to 15 million purified oocysts per mouse. For in vitro cultivation, purified oocysts were pretreated in a low pH (2.5- 3) 0.5% trypsin solution for 20 min, resuspended in supplemented RPMI-1640 containing glucose 0.1 g (5.55 mM), sodium bicarbonate 0.3 g, bovine bile 0.02 g, folic acid 25 μg, 4-aminobenzoic acid 100 μg, calcium pantothenate 50 μg, ascorbic acid 875 μg, penicillin G 10,000 U and streptomycin 0.01 g per 100 ml, and 1% fetal bovine serum (pH 7.4 before filtration), and used to inoculate confluent monolayers of the human adenocarcinoma cell line HCT-8. Incubation was in a candle jar at 37 C. We tested numerous supplements to RPMI-1640, different pHs, and atmospheric conditions and found the parameters described above produced the greatest parasite numbers in vitro. We obtained significantly superior growth of C. parvum grown in HCT-8 cells using the conditions described above than in culture conditions described previously.
Details
- Title
- Simplified methods for obtaining purified oocysts from mice and for growing Cryptosporidium parvum in vitro
- Authors/Creators
- B.P. Meloni (Author/Creator)R.C.A. Thompson (Author/Creator)
- Publication Details
- The Journal of Parasitology, Vol.82(5), pp.757-762
- Publisher
- American Society of Parasitology
- Identifiers
- 991005543055407891
- Murdoch Affiliation
- School of Veterinary Studies
- Language
- English
- Resource Type
- Journal article
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- Citation topics
- 1 Clinical & Life Sciences
- 1.246 Diarrheal Diseases
- 1.246.985 Cryptosporidium
- Web Of Science research areas
- Parasitology
- ESI research areas
- Microbiology