Hi Afsana Kajee,
I infected Thp1 cells with Lactobacillus acidophilus and Bacillus clausii. These cells as such mostly not intake these bacteria, however needs to be differentiated into macrophage before being studied for its uptake capacity. Differentiate Thp1 monocytic cell lines into macrophage by treating it with PMA for 24 hours, then resting for 72 hours. Then treat the macrophage with C. neoformans in different multiplicity of infection (MOI). For bacterial uptake, optimal time of treatment is 2 hours. But C. neoformans it may take longer time, you have to standardize it. You can refer my article http://link.springer.com/article/10.1007/s12602-016-9211-4 for detailed protocol and methods used.
Hi Afsana Kajee,
I infected Thp1 cells with Lactobacillus acidophilus and Bacillus clausii. These cells as such mostly not intake these bacteria, however needs to be differentiated into macrophage before being studied for its uptake capacity. Differentiate Thp1 monocytic cell lines into macrophage by treating it with PMA for 24 hours, then resting for 72 hours. Then treat the macrophage with C. neoformans in different multiplicity of infection (MOI). For bacterial uptake, optimal time of treatment is 2 hours. But C. neoformans it may take longer time, you have to standardize it. You can refer my article http://link.springer.com/article/10.1007/s12602-016-9211-4 for detailed protocol and methods used.
Hi Biswaranjan Pradhan
Thank you for the help, I will refer to your article.
All the best :-)
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