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Effect of select medium supplements on in vitro development of Cryptosporidium andersoni in HCT-8 cells

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Abstract

We evaluated the effect of fetal calf serum (FCS), glucose, ascorbic acid, calcium pantothenate, folic acid, and insulin on the growth of Cryptosporidium andersoni in human colon tumor (HCT-8) cells. After being incubated for 48 h, the proliferation of parasites was determined by real-time polymerase chain reaction (PCR) assay, and the development of C. andersoni was observed by transmission electron microscopy (TEM). Ten percent FCS was the best concentration for C. andersoni culture. Glucose, ascorbic acid, and insulin had a significant effect on the growth of C. andersoni when added into 10% FCS RPMI 1640. Calcium pantothenate had no significant effect and folic acid had the inhibited effect. We also observed the stages of trophozoite, macrogamont, microgamont, type I meront, type II meront, and sporozoite of C. andersoni in HCT-8 cells by TEM. Our results indicated that the best medium for C. andersoni was 10% FCS RPMI 1640 medium containing 50 mM glucose, 50 μg/ml ascorbic acid, and 0.3 U/ml insulin. Real-time PCR could provide a quick and precise technique to determine the proliferation of parasites. Cultivation of C. andersoni in HCT-8 cells will facilitate the study of interactions between parasites and host cells as well as provide a reliable system for evaluating anticryptosporidial compound efficacy.

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Acknowledgments

The authors would like to thank Xiao, L. at the Division of Parasitic Diseases, National Center for Zoonotic, Vector-Borne, and Enteric Diseases, Centers for Disease Control and Prevention, Atlanta, USA for the help with the identification of Cryptosporidium species and Martin Kváč at the Institute of Parasitology, Academy of Sciences of the Czech Republic for identification of the stages of C. andersoni in HCT-8 cells by TEM. This study was supported by the Laboratory of Parasite and Vector Biology, MOH (No. WSBKFKT200701), the National Resource Platform Project from Ministry of Science and Technology (No. 2005DKA 21104), the Sci-Tech Innovation Team of Jiangsu University (No. 2008-018-02), the National Special Program for Scientific Research of Public Health (No. 200802012) and the National Key Program for Infectious Diseases of China (No. 2008ZX1004, No. 2009ZX10602). The execution of the experiments described in this article complied with all applicable laws of the People’s Republic of China.

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Authors and Affiliations

  1. School of Medical Science and Laboratory Medicine, Jiangsu University, 212013, Zhenjiang, People’s Republic of China

    Liang Wu, Sheng-xia Chen, Xu-gan Jiang, Zhao-xi Lu, Guo-hua Tu & Xing-li Fu

  2. National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Laboratory of Parasite and Vector Biology, MOH, 200025, Shanghai, People’s Republic of China

    Yu-juan Shen & Jian-ping Cao

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  1. Liang Wu
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  2. Sheng-xia Chen
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  3. Xu-gan Jiang
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  7. Xing-li Fu
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Correspondence to Sheng-xia Chen.

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Wu, L., Chen, Sx., Jiang, Xg. et al. Effect of select medium supplements on in vitro development of Cryptosporidium andersoni in HCT-8 cells. Parasitol Res 105, 1419–1424 (2009). https://doi.org/10.1007/s00436-009-1576-9

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  • DOI: https://doi.org/10.1007/s00436-009-1576-9

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