The invention puts forward a universal cancer organoid in-vitro culture medium, which consists of a DMEM/F-12K (Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12K) basic culture medium and an adding factor, wherein the adding factor consists of HEPES (2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid), L-Glutamine, EGF (epidermal growth factor), Noggin, FGF (fibroblast growth factor)-10, A83-01 and Y27632. According to the culture medium of the embodiment of the invention, the success rate of organoid passage culture can be greatly improved, in addition, the universal culture of cancer samples, including gastric carcinoma, rectal carcinoma, lung carcinoma and the like, can be realized, culture cost is greatly lowered, the long-term culture of the organoid can be realized, and abiobank is established.