The literature reveals array of assay including assay using tetrazolium dyes, using mix of sybr green in Triton, counting using cytometer, etc.
I would appreciate if the expert working in said field could assist me with the most efficient way along with the protocols to assess the growth inhibition of said culture system.
Thanks
First of all, I would recommend to use amastigote forms (either axenic or in macrophages) instead of promastigote ones. The amastigote form is the target of chemotherapy and the promastigote forms only represent them partially. The most simple protocol is using Alamar blue (resazurin) as an endpoint. You can contact Marcel Kaiser ([email protected]) he has established protocols which also work for promastigotes.
First of all, I would recommend to use amastigote forms (either axenic or in macrophages) instead of promastigote ones. The amastigote form is the target of chemotherapy and the promastigote forms only represent them partially. The most simple protocol is using Alamar blue (resazurin) as an endpoint. You can contact Marcel Kaiser ([email protected]) he has established protocols which also work for promastigotes.
I would agree with Reto re amastigotes. We have published a number of protocols over the past few years that use a variety of readouts including bactiterglo and high content imaging. I would use caution in the design and analysis of the data. It is not enough to have compounds that slow the growth or stop parasites growing. We would suggest that the compounds have to kill the parasites for this to translate effectively into animal models. One of the major reasons for mismatches between axenic and cell based assays is that the axenic unless set up carefully reports growth slowing and cytostatic compounds as active whereas the high content screen reports cell killing. Suggest reading doi 10.1128/AAC.02398-12 . All the best
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