Rapid Dissociation of Tumor Tissue to Gain Enhanced Single-Cell Tumor Insights

To begin tumor digestion, pour the sample into a 60-millimeter tissue culture dish placed on ice. Pipette 420 microliters of media from the dish to a 1.5-milliliter microcentrifuge tube. Using tweezers, transfer the sample tissue to the tube containing media.

Cut up the sample with clean scissors to pieces that are two to three millimeters in diameter. Add appropriate volumes of digestion enzymes and media to the sample. Vortex the tube for five seconds, and incubate it in a thermal mixer positioned vertically on its side.

Place a 50-micrometer cell filter on top of a new 15-milliliter conical tube. And prime the filter with one milliliter of fresh RPMI media. Using a 1, 000-microliter lower tension wide pipette tip, load the sample to the filter and add the media.

Then, with the back of a syringe plunger, grind and push the sample on the filter in a twisting motion. Wash the filter with five milliliters of media and any remaining media from the dish that contain the sample.