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Oncogenic driver supersedes BM preparation as the critical determinant of leukemic outcome: a one day protocol for BM harvest, infection and transplantation

Bone Marrow Transplantation volume 48, pages 1003–1005 (2013)Cite this article

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Infection of murine BM, followed by transplantation of the infected cells into lethally irradiated recipient mice is a critical assay for understanding how oncogenes alter leukemia progression and hematopoietic lineage decisions. Conventional wisdom suggests that donor mice should be treated with 5-fluorouracil (5FU) and harvested BM cells cultured in the presence of cytokines for up to 5 days to enrich for the stem cells for these experiments to be successful. However, we show that regardless of how donor BM is prepared, the leukemic phenotypes and disease progression is dependent only on the oncogenic driver expressed in the infected cells.

Our laboratory has been interested in the role of oncogenic combinations in the initiation, progression and maintenance of leukemia.1, 2, 3 To this end, we have adopted a BM infection and transplantation model as a rapid, reliable and flexible model system. Penetrance, latency and cell surface phenotype of leukemias differ depending on the oncogenic combinations being tested. Published protocols suggest that mice should be treated with 150 mg/kg of 5FU for at least 5 days followed by harvesting cells from bones and then culturing the cells in the presence of SCF, IL-3 and IL-6 for a minimum of 2 days.4 At the end of the culture period, the cells are infected with viral supernatant by centrifugation in the presence of polybrene. Some protocols call for multiple rounds of infection over multiple days. Overall, this protocol is time consuming and costly.

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References

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Acknowledgements

I would like to thank Harold Varmus for his support and discussion during this work. Also the members of the Varmus laboratory during my time at MSKCC, especially G Sanchez, A Giannakou, R DeMatteo and MA Melnick, for expert handling of the mouse colony, including genotyping. National Institutes of Health, Molecular Targets COBRE, 8P20GM103482-10 (to LJB), Wendy Will Case Cancer Fund, GB220413 (to LJB.), Kosair Pediatric Cancer Research Program award (to LJB)

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  1. Division of Hematology and Oncology, Department of Medicine, James Graham Brown Cancer Center, University of Louisville, Louisville, KY, USA

    L J Beverly

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  1. L J Beverly
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Correspondence to L J Beverly.

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Beverly, L. Oncogenic driver supersedes BM preparation as the critical determinant of leukemic outcome: a one day protocol for BM harvest, infection and transplantation. Bone Marrow Transplant 48, 1003–1005 (2013). https://doi.org/10.1038/bmt.2012.273

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