I will be isolating T cells from blood samples and culturing them in a T-75 flask. What would be the required blood volume needed? Or rather, how much blood would I need to have enough of cells to culture in a T-75?
In my experience, there are around 4 to 5 million T cells in 6 mL of peripheral venous blood of healthy adult donors (the reference range for CD3+ T cells in circulation is between 0.67 - 2.38 x 10^6/mL). The question is, after you isolate the peripheral blood mononuclear cells, how do you plan to isolate the T cells? The isolation efficiency depends on the method. Flow cytometry-based or magnetic-based sorting may yield around 90-95% efficiency. I would recommend to culture around 4 million T cells in one 75 cm2 tissue culture flask. However, this is dependent on your assay, what kind of differentiation/activation/expansion protocol are you using.
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