How can I check PBMCs viability after treatment with a substance(...

protocol Protocol: In vitro Culture of Human PBMCs
How can I check PBMCs viability after treatment with a substance( carvacrol) ? I have tried MTT assay but I think that is not enough! So, i want to try with Trypan blue 0,4%. I have my cells in 96 well plate. After 24h incubation with carvacrol how can I check viability with trypan blue?
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Santosh K Panda Author Answered Apr 26, 2016

Infectious Disease Biology, Institute of Life Sciences, India

Hi
Loanna
My boss Dr. Ravindran answer this to a previous query. here it is, Cells in PBMC constituting Monocytes and Lymphocytes are terminally differentiated cells normally. Monocytes do not divide and can only be differentiated to macrophages by adding cytokines into culture. Lymphocytes also will not divide unless activated by a variety of means. Under normal culture conditions cells will die over time.
hope this will be helpful.
thanks
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Ioanna Eleftheriadou Answered Apr 25, 2016

Aristotle University of Thessaloniki

Thank you very much. I have one more question: Do the PBMCs proliferate without mitogen?I seed 150000cells/well and incubate with a plant extract. 20 hours later, i count more cells. Which is the proliferation rate of PBMCs?
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Santosh K Panda Author Answered Apr 25, 2016

Infectious Disease Biology, Institute of Life Sciences, India

Hi Loanna
You can remove some cells from the plate and can use trypans blue. otherwise you can also stain them by DAPI and check by FACS.
thanks
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