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Year : 2009  |  Volume : 27  |  Issue : 1  |  Page : 35-39

In vitro culture of various species of microsporidia causing keratitis: Evaluation of three immortalized cell lines


1 Jhaveri Microbiology Center, Brien A Holden Eye Research Centre, Hyderabad Eye Research Foundation, L. V. Prasad Eye Institute, Hyderabad, India
2 Ocular Microbiology Service, L. V. Prasad Eye Institute, Bhubaneswar, India

Correspondence Address:
S Sharma
Ocular Microbiology Service, L. V. Prasad Eye Institute, Bhubaneswar
India
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Source of Support: None, Conflict of Interest: None


PMID: 19172057

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Being intracellular parasites, microsporidia can only be propagated in cell culture systems. This study evaluated three cell lines to determine the most suitable host-parasite In vitro system. Confluent monolayers of vero, SIRC, and HeLa cell lines, grown in 24-well tissue culture plates, were inoculated with varying concentrations (1 10 4 to 1 10 8 spores/mL) of Vittaforma corneae, Encephalitozoon hellem, Encephalitozoon cuniculi, and Encephalitozoon intestinalis spores. Growth was compared quantitatively at weekly intervals. Encephalitozoon species showed the highest amount of growth when cultured in vero cell line, while there was no significant difference in their growth in SIRC and HeLa cell lines. In comparison, V. corneae showed the highest growth in SIRC cells, followed by vero cells. The analytical sensitivity was found to be 1 10 4 spores/mL for vero cell line compared to 1 10 5 spores/mL for SIRC cell line and 1 10 7 spores/mL for HeLa cell line. HeLa cells also showed rapid disruption of cells, and the spores could not be easily distinguished from cell debris. This is the first report of the comparison of vero, SIRC, and HeLa for the propagation of microsporidial spores. Vero cell line was found to be more sensitive than SIRC and HeLa cells, and we believe that the inclusion of vero cell line in the routine culture protocols of ocular parasitology laboratories would result in a significant increase in the diagnostic yield.






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