| abstract |
The invention discloses a novel method for culturing colorectal cancer organoids, comprising the following steps: a cell processing step: removing a part of tumor tissue, and putting the surgically removed sample into tissue protection solution and transporting it to a laboratory for ultra-cleaning In Taichung, the cancer tissues were repeatedly washed with tissue washing solution, the remaining cancer tissues were cut into 1xmm2 pieces with sterile scissors, the digestion solution was added, the digestion solution was shaken at 37°C for 1-3 hours, centrifuged, the supernatant was discarded, and normal saline was added. Use a cell strainer to filter out the tissue pieces. If there are too many red blood cells, add red blood cell lysate to lyse the red blood cells. Double-antibody CD44 and EpCAM staining are performed on the digested single cells. Separation and culture steps: The isolated cells are incubated with PI to exclude dead cells. In the present invention, the CD44-positive cancer stem cells and the CD44-negative non-stem cells are separated and cultured, and then the CD44-positive stem cells are expanded and cultured to improve the success rate of tumor organoid culture. |