Experimental Competitive Bone Marrow Transplant Assays

Hematopoietic stem cells have the ability to produce all blood cells. When hematological malignancies occur, transplant of compatible blood or bone marrow cells from a healthy donor to the patient is an efficient solution to restore normal hematopoiesis. Bone marrow transplant in a mouse model is often used to study HSC function and capacity to repopulate an irradiated recipient. This protocol details the different steps of a competitive bone marrow transplant experiment, beginning with total body irradiation of the recipient mice; preparation and administration of the donor and competitor bone marrow samples; peripheral blood analysis to follow reconstitution posttransplant; and finally, the analysis of recipient bone marrow and secondary transplants to evaluate long-term HSC function. Different formulas used to establish transplant efficiency are explained. All the steps are discussed in detail, including tips, variations, and alternative procedures with their advantages and disadvantages.