I am trying to induce toxoplasma cyst formation (both RH and ME49) strains by altering the pH to 8.1 (DMEM, 1%FCS, 1% pen-strep), at 37oC, without CO2. However, the host cells weren't healthy and dying after 24 hours of incubation in condition without C02, any suggestion in improving this? As I understand that the change in media pH is the most common way to induce cyst formation in vitro. Thanks.
Long - term culturing of Toxoplasma gondii cysts has been accomplished in vitro in association with murine astrocytes and intermittent additions of gamma interferon to the media. Phase - contrast microscopy was used to follow the stages of cyst development., and electron microscopy confirmed the presence of morphologic characteristics of T. gondii cysts. T. gondii cysts formed in vitro had a single trilaminar membrane during both intracellular and extracellular existence and contained amorphous electron - dense material either throughout the cyst or in a unform layer under the trilaminar membrane........ in vitro induction of nonspecific resistance in macrophages by specifically sensitized lymphocytes. For more details consult https://www.ncbi.nlm.nih.gov
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