Efficient derivation of immortalized, isogenic cell lines from genetically defined murine hepatoblastomas

Hepatoblastoma (HB), the most common childhood liver cancer and the simplest of human neoplasms molecularly, commonly deregulates Wnt/β-catenin (B), Hippo/YAP (Y) and/or NFE2/NRF2 (N) signaling. Murine HBs that closely resemble human tumors can be generated by over-expressing pairwise or triple combination of mutant forms of B,Y and N. However, research is hampered by a paucity of established cell lines from either species. We show here that immortalized cell lines can be routinely established from murine HBs that over-express oncogenic B + Y and B + Y + N if the Cdkn2a tumor suppressor locus is concurrently mutated as also commonly occurs in human HBs. Eight BY and 3 BYN cell lines are characterized here, 10 of which can be regrown as subcutaneous and metastatic lung tumors in the immune-competent mice from which they originated. During maintenance as hypoxic spheroids in vitro , tumor cells express endothelial cell markers. In vivo , similar cells appear adjacent to blood vessels. The ability to routinely generate isogenic HB cell lines with defined oncogenic drivers should facilitate studies that are best performed in vitro . The approach may also be useful for deriving HB cell lines associated with less common molecular drivers and from human tumors. • Few murine or human hepatoblastoma cell lines currently exist. • We describe 11 murine lines driven by 3 of the most common HB-associated oncogenes. • The cell lines can be readily propagated in immuno-competent mice. • The method of cell line derivation is generalizable.