Korean J Parasitol.  2004 Mar;42(1):27-34. 10.3347/kjp.2004.42.1.27.

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

Affiliations
  • 1Department of Parasitology and Tropical Medicine, Seoul National University College of Medicine, and Institute of Endemic Diseases, Seoul National University Medical Research Center, Seoul 110-799, Republic of Korea.
  • 2Department of Therapeutic Radiology, Chungbuk National University College of Medicine, Cheongju 361-763, Republic of Korea.
  • 3Department of Parasitology, College of Medicine, Konkuk University, Chungju 380-701, Republic of Korea.

Abstract

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.

Keyword

Cryptosporidium muris; human stomach adenocarcinoma (AGS) cell; in vitro culture; medium supplement

MeSH Terms

Adenocarcinoma
Animals
Cell Line, Tumor
Cryptosporidium/*growth & development
Culture Media
Human
Hydrogen-Ion Concentration
Stomach/*parasitology
Stomach Neoplasms
Support, Non-U.S. Gov't
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