Authors

Discussion

INTRODUCTION: Recently there has been the development of several culture platforms for the propagation of Cryptosporidium. The simplest of these being through the infection of mammalian COLO-680N monolayers.

AIMS: We aimed to optimise and adapt this culture system for the purpose of assessing the efficacy and selectivity of therapeutic agents against Cryptosporidium.

METHODS: To do this we established 5mL flask culture systems and monitored the number of motile parasites released from COLO-680N cells. From this, we have subsequently reduced the culture volume to increase parasite growth throughput. Recently we have evaluated the potential use of the cytochemical Alamar Blue to detect Cryptosporidium growth using this platform.

RESULTS: Initial results show evaluating the number of motile merezoites and meronts in flask and plate cultures is an effective measure for gauging selective toxicity of compounds towards Cryptosporidium. Alamar blue was not effective for discriminating merezoite growth, but shows potential in host cell protection from infection.

DISCUSSION: Straightforward culture of Cryptosporidium has permitted the development of merezoite and meront inhibitor assays, which are likely to lead to the identification of novel therapeutic agents in the near future.