| abstract |
The present invention relates to the field of tumor organoid culture. Specifically, it relates to a serum-free special culture medium and a culture method for pancreatic cancer organoids. The medium is a serum-free special medium without the addition of recombinant proteins, and consists of a basal medium and additives. The basal medium is a medium containing 1% penicillin, 1% HEPES buffer, and 1% GlutaMax solution. DMEM/F12 medium. Culture supplements include growth factors EGF, FGF2, FGF7, FGF10, Neuregulin1, insulin, hydrocortisone, small molecule inhibitors A83‑01, Y‑27632, SB202190, B27 supplement. Patient-derived pancreatic cancer samples were digested with collagenase, resuspended in Cultrex TM growth factor reduced BME type2, added to a culture plate, solidified at 37 degrees for half an hour, added with the above medium, placed in a cell incubator, and the medium was changed every 4 days , the desired pancreatic cancer organoids can be obtained in one week. The culture medium of the present invention can support the growth of pancreatic cancer cells in vitro, present a typical spherical or irregular mass tumor organoid shape, and provide a good research sample for subsequent further research. |