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Research Article

CD40 ligand is necessary and sufficient to support primary diffuse large B-cell lymphoma cells in culture: a tool for in vitro preclinical studies with primary B-cell malignancies

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Pages 1390-1398 | Received 02 Sep 2011, Accepted 29 Dec 2011, Published online: 03 Feb 2012
 

Abstract

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.

Acknowledgements

The authors would like to thank all the dog owners who allowed their pets to participate in this study, as well as Drs. Antonella Borgatti and Michael Henson (University of Minnesota, Minneapolis/St. Paul, MN), Dr. Kristine Burgess (Tufts Cummings School of Veterinary Medicine, North Grafton, MA), Dr. Gretchen Gerber (Country Care Pet Care, Washburn, WI) and Dr. Janice Baserga (Scarborough Animal Hospital, Scarborough, ME) for assistance with collecting biopsy samples. We wish to acknowledge the assistance of Dr. Aaron Sarver (University of Minnesota) for microarray data analysis and also thank Drs. Aaron Sarver, Jeffrey Miller, Michael Verneris (University of Minnesota) and Douglas Thamm (Colorado State University, Fort Collins, CO) for critical review of the manuscript and helpful discussions.

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).

Potential conflict of interest

Disclosure forms provided by the authors are available with the full text of this article at www.informahealthcare.com/lal.

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