Parasites, Hosts and Diseases

  • 제42권1호
  • /
  • Pages.27-34
  • /
  • 2004
  • /
  • 2982-5164(pISSN)
  • /
  • 1738-0006(eISSN)
대한기생충학·열대의학회 (The Korea Society for Parasitology and Tropical Medicine)
권호 표지

In vitro culture of Cryptosporidium muris in a human stomach adenocarcinoma cell line

  • Choi, Min-Ho (Department of Parasitology and Tropical Medicine, and Institute of Endemic Diseases, Seoul National University College of Medicine and Medical Research Center) ;
  • Hong, Sung-Tae (Department of Parasitology and Tropical Medicine, and Institute of Endemic Diseases, Seoul National University College of Medicine and Medical Research Center) ;
  • Chai, Jong-Yil (Department of Parasitology and Tropical Medicine, and Institute of Endemic Diseases, Seoul National University College of Medicine and Medical Research Center) ;
  • Park, Woo-Yoon (Department of Therapeutic Radiology, Chungbuk National University College of Medicine) ;
  • Yu, Jae-Ran (Department of Parasitology, College of Medicine, Konkuk University)
  • 발행 : 2004.03.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

We investigated the optimal culture conditions for Cryptosporidium muris in a human stomach adenocarcinoma (AGS) cell line by determining the effects of medium pH and of selected supplements on the development of C. muris. The optimum pH of the culture medium required for the development of C. muris was determined to be 6.6. The number of parasites significantly increased during cultivation for 72 hr (p < 0.05) at this level. On the other hand, numbers decreased linearly after 24 hr of incubation at pH 7.5. When cultured in different concentrations of serum, C. muris in media containing 5% FBS induced 4-7 times more parasites than in 1% or 10% serum. Of the six medium supplements examined, only 1 mM pyruvate enhanced the number of C. muris in vitro. Transmission electron microscopic observation showed the developmental stages of C. muris in the cytoplasm of the cells, not in an extracytoplasmic location. The growth of C. muris in AGS cells provides a means of investigating its biological characteristics and of testing its response to therapeutic agents. However, a more optimized culture system is needed for the recovery of oocysts on a large scale in vitro.

키워드

참고문헌

  1. Current WL, Garcia LS (1991) Cryptosporidiosis. Clin Microbiol Rev 4: 325-358 https://doi.org/10.1128/CMR.4.3.325
  2. Fayer R, Leek RG (1984) The effects of reducing conditions, medium, pH, temperature, and time on in vitro excystation of Cryptosporidium. J Protozool 31: 567-569 https://doi.org/10.1111/j.1550-7408.1984.tb05504.x
  3. Gatei W, Ashford RW, Beeching NJ, Kamwati SK, Greensill J, Hart CA (2002) Cryptosporidium muris infection in an HIV-infected adult, Kenya. Emerg Infect Dis 8: 204-206
  4. Gut J, Petersen C, Nelson R, Leech J (1991) Cryptosporidium parvum: in vitro cultivation in Madin-Darby canine kidney cells. J Protozool 38: 72S-73S
  5. Hamer DH, Ward H, Tzipori S, Pereira ME, Alroy JP, Keusch GT (1994) Attachment of Cryptosporidium parvum sporozoites to MDCK cells in vitro. Infect Immun 62: 2208-2213
  6. Hijjawi NS, Meloni BP, Morgan UM, Thompson RC (2001) Complete development and long-term maintenance of Cryptosporidium parvum human and cattle genotypes in cell culture. Int J Parasitol 31: 1048-1055 https://doi.org/10.1016/S0020-7519(01)00212-0
  7. Hijjawi NS, Meloni BP, Ryan UM, Olson ME, Thompson RC (2002) Successful in vitro cultivation of Cryptosporidium andersoni: evidence for the existence of novel extracellular stages in the life cycle and implications for the classification of Cryptosporidium. Int J Parasitol 32: 1719-1726
  8. Iseki M, Maekawa T, Moriya K, Uni S, Takada S (1989) Infectivity of Cryptosporidium muris (strain RN 66) in various laboratory animals. Parasitol Res 75: 218-222 https://doi.org/10.1007/BF00931279
  9. Katsumata T, Hosea D, Ranuh IG, Uga S, Yanagi T, Kohno S (2000) Short report: possible Cryptosporidium muris infection in humans. Am J Trop Med Hyg 62: 70-72
  10. Meloni BP, Thompson RC (1996) Simplified methods for obtaining purified oocysts from mice and for growing Cryptosporidium parvum in vitro. J Parasitol 82: 757-762 https://doi.org/10.2307/3283888
  11. Petry F, Robinson HA, McDonald V (1995) Murine infection model for maintenance and amplification of Cryptosporidium parvum oocysts. J Clin Microbiol 33: 1922-1924
  12. Tzipori S, Ward H (2002) Cryptosporidiosis: biology, pathogenesis and disease. Microbes Infect 4: 1047-1058
  13. Uni S, Iseki M, Maekawa T, Moriya K, Takada S (1987) Ultrastructure of Cryptosporidium muris (strain RN 66) parasitizing the murine stomach. Parasitol Res 74: 123-132 https://doi.org/10.1007/BF00536023
  14. Upton SJ, Tilley M (1992) Effect of select media supplements on motility and development of Eimeria nieschulzi in vitro. J Parasitol 78: 329-333 https://doi.org/10.2307/3283483
  15. Upton SJ, Tilley M, Brillhart DB (1995) Effects of select medium supplements on in vitro development of Cryptosporidium parvum in HCT-8 cells. J Clin Microbiol 33: 371-375
  16. Widmer G, Corey EA, Stein B, Griffiths JK, Tzipori S (2000) Host cell apoptosis impairs Cryptosporidium parvum development in vitro. J Parasitol 86: 922-928
  17. Yu JR, Choi SD (2000) The effect of microfilament inhibitor on the Cryptosporidium infection in vitro. Korean J Parasitol 38: 257-261
  18. Yu JR, Choi SD, Kim YW (2000) In vitro infection of Cryptosporidium parvum to four different cell lines. Korean J Parasitol 38: 59-64