The application discloses a preparation method and application of large-size
human liver organoids, and belongs to the technical field of
cell culture. In the application, 2D culture mature human primary liver parenchymal cells with different
cell amounts are resuspended by using a
hepatocyte maturation medium HIM, and are added into a U-shaped bottom 96-hole plate for low-
oxygen static culture. After the cells are aggregated, the cells are cultured in normal
oxygen and are shaken, so that large-size
human liver organoids with different
cell contents are prepared. The method does not need
biological materials, and can induce human primary liver parenchymal cells to self-assemble and construct functional liver organoids with high uniformity in scale through specific culture conditions. The method solves the problems of large batch difference, high heterogeneity and dedifferentiation in the prior art, and can be widely applied to
drug hepatotoxicity testing, in-vitro modeling of liver diseases and development of a bioartificial liver
system.