Successful Establishment of an Orthotopic Hepatoblastoma In Vivo Model in NOD/LtSz-scid IL2Rγnull Mice

Abstract

Investigation of hepatoblastoma in experimental conditions contributes relevantly to a detailed understanding of tumor biology and the investigation of new treatment approaches. Most systematical analyses currently use subcutaneous xenografts. We established a reproducible intrahepatic model with the hepatoblastoma-cell lines HuH6 and HepT1. The cells were stably transfected with a plasmid vector encoding for Gaussia luciferase. HuH6 and HepT1 were injected intrasplenically in NOD/LtSz-scid IL2Rγnull mice. Mice were splenectomized in order to avoid intrasplenical tumor growth. Multifocal intrahepatic tumor growth was observed in 85% (11/13) of HuH6 tumors and 55% (5/9) of HepT1 tumors. Serum Alpha-fetoprotein and Gaussia luciferase increased 5 weeks after tumor-cell inoculation. Tumors were detected by MRI at this time point. Immunhistochemical analysis such as vascularity (CD31), proliferation index (Ki-67), cytokeratin 7 and distribution of β -catenin in intrahepatic tumors were different to subcutaneous tumors. We established a reproducible xenograft model for intrahepatic hepatoblastoma growth with a high tumor incidence. Monitoring of tumor cell viability was optimized by measuring GLuc. This model enables further experimental investigations of HB in a more physiological milieu as emphasized by the β-catenin distribution.

Novel patient-derived xenograft and cell line models for therapeutic testing of pediatric liver cancer

“…These models represent a certain degree of tumor heterogeneity, but their utility is limited by the fact that the resulting tumors are encapsulated (rather than invasive), and so far subcutaneous HB have not been shown to form metastases. Another type of tumor model, often used for in vivo drug studies, involves the injection of human HB cell lines (rather than primary tumor cells) into the subcutaneous tissue or splenic capsule of immune-deficient mice [13]. These models are also limited, however, because unlike the primary tumors, the cell lines are usually monoclonal and have been selected for their ability to grow in tissue culture.…”

Targeting EpCAM (CD326) for immunotherapy in hepatoblastoma

“…Xenografts derived from HUH6 cells with stable expression of Gaussia luciferase (GLuc) (pCMV-GLuc, NEB) were developed, as described previously 41 . Spare tumor tissue was sliced in culture media with two scalpels as longitudinal sections of 2–3 mm thickness.…”

“…Xenografts were derived from HUH6 and HepT1 cells, as described elsewhere 41 . Tumor specimen from a patient suffering of HB and receiving CDDP-based chemotherapy was obtained after informal consent.…”

Animal models of extracranial pediatric solid tumors

“…An intrasplenic injection of hepatoblastoma cells was administered into a NOD/LtSz-scid IL2rγ null mouse model. A splenectomy was then performed, which led to a high tumor incidence in the liver (19).…”