bulk removal of dead cells
ZBIGNIEW DARZYNKIEWICZ (darzynk@nymc.edu)
Mon, 23 Jun 1997 10:21:26 -0500
Efficient removal of dead cells (necrotic, late apoptotic- i.e. at
necrotic stage of apoptosis, cell debris, mechanically broken cells,
cell nuclei) can be obtained by suspending the cell pellet (e.g.
obtained from mechanically disaggregated tumors) in PBS solution
containing 200 ug/ml of DNase I, incubating them for 15 min, then
adding trypsin to a final concentration 0.25% and incubating for an
additional 15 - 30 min. The trypsin is then neutralized by adding
medium with 10% serum, the cells are centrifuged and resuspended in
serum or albumin containing medium. Cell viability (trypan blue or PI
assay) is then close to 100% (see page 20, Meth. Cell Biol. Vol 41,
1994).
Zbigniew Darzynkiewicz
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