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Comparison of Reproduction Densities in Different Liquid Media of Trypanosoma cruzi and Cryopreservation.

OBJECTIVE
This study aims to determine the optimum liquid medium for the reproduction of Trypanosoma cruzi strains and provide cryopreservation.


METHODS
The reproduction density of T. cruzi strain was evaluated in the following five different commercial liquid culture media: RPMI 1640, Medium 199 (M199), Schneider's İnsect Medium (SİM), Nutrient Broth (NB), Brain Heart İnfusion Broth (BHİB). Cultures were monitored on every other day for a period of 24 days. Cryopreservation of T. cruzi was also performed and viability was tested after six months.


RESULTS
Epimastigotes of T. cruzi were not found to be produced in NB and BHIB media. Significant difference was not observed among the reproduction potential of RPMI-1640, M199, and SIM after evaluating the data for the first 10 days. Between days 12 and 24, RPMI-1640 was found to be the best reproduction medium. From the 18th day onwards, parasites transformed amastigotes. On the 24th day, the highest level of amastigote amount was observed, and reproduction was determined to have stopped. As a result of cryopreservation, it was determined that the survival of T.cruzi continued after six months..


CONCLUSION
Thus, the selection of RPMI-1640 medium, followed by M199 and SIM media would be appropriate when studying T. cruzi epimastigotes. Studies using epimastigotes should be planned for up to 18 days and for those using amastigotes, it would be appropriate to plan the studies after the 18th day. Moreover, T. cruzi can be cryopreserved with 15% DMSO and stored for a long time in liquid nitrogen.