Established cell lines are utilized extensively to study tumor biology and preclinical therapeutic development. However, they may not accurately recapitulate the heterogeneity of their corresponding primary disease. B-cell tumor cells are especially difficult to maintain under conventional culture conditions, limiting access to samples that faithfully represent this disease for preclinical studies. Here, we used primary canine diffuse large B-cell lymphoma to establish a culture system that reliably supports the growth of these cells. CD40 ligand, either expressed by feeder cells or provided as a soluble two-trimeric form, was sufficient to support primary lymphoma cells in vitro. The tumor cells retained their original phenotype, clonality and known karyotypic abnormalities after extended expansion in culture. Finally, we illustrate the utility of the feeder cell-free culture system for comparable assessment of cytotoxicity using dog and human B-cell malignancies. We conclude that this system has broad applications for in vitro preclinical development for B-cell malignancies.
Bibliographical noteFunding Information:
This work was supported by MAF First Award Grant D12CA-302 (DI), AKC Canine Health Foundation grant 615 (JFM and MB), NIH grant R01CA112211 (MB), AKC Canine Health Foundation grant 1113 (TDOB and JFM), NIH grant P30CA077598 (NCI Core Support Grant for the Masonic Cancer Center), and by philanthropic funds from the University of Minnesota Animal Cancer Care and Research Program, the Starlight Fund, and the Land of PureGold Foundation, Inc (JFM).
- Diffuse large B-cell lymphoma
- canine model
- cytotoxicity assay
- primary cell culture