||Depletion of Dead Cells and Debris from Tumor Digestion
||~300-500 mg of tumor was enzymatically digested.
Cells were washed and resuspended in Annexin binding buffer.
Cells were washed once more then resuspending in 200 uL of 1:50 dilution of Annexin-biotin for 20 minutes at room temperature.
Cells were washed once with binding buffer then incubated with magnetic streptavidin beads at room temperature for 20 minutes
Cell suspension was then incubated on magnet and the negative fraction was retained.
||My tumor digestions contained a lot of dead cells and debris that interfere with downstream stimulation and other applications. This was highly efficient for removing dead cells and debris.
||Results varied slightly by tumor but typically 70-95% of dead/cells debris were removed as assessed by trypan blue staining on a hemacytometer.
Subsequent staining we do results in more dead cells, but representative FSC/SSC is shown in image. Really nice purification of tumor infiltrating lymphocytes.