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Isolate CD4+ T Cells - Whole Blood, Buffy Coat, MNC, Bone ...

https://www.lifetechnologies.com/...buffy.../isolate-cd4-t-cells.html

Buffy coat is 8-10 times more concentrated than whole blood with regard to ... MNC 1ml (1 x 107 cells); washed whole blood 1 ml; washed and diluted buffy coat 1 ml ... To obtain residual cells, wash the beads 2-3 times in 500 μl Buffer 2 and ... Wash detached cells thoroughly by resuspending the cells in a total volume of 10 ..

Dear Oriana,

in my experiments,  a buffy coat (180 ml) yielded 2.5x10E8 PBMC on average (range 1.2 - 6 x 10E8, depending on the healthy blood donor) after ficol density gradient centrifugation.

This complies with Ram Nath's answer.

Good luck,

Sabine

Hi Oriana,

You can use Ficoll-histopaque-1077 for better isolation of lymphocytes, After isolation dissolve the cell pellet in few microliters (50 to 100) of PBS or RPMI medium and count the number of cells using cell counting chamber. based on the cell count u can dissolve the pellet as you required. Generally, 1-2 ml will give sufficient number of cells.

Thank you all!