Abstract
Competitive bone marrow transplantation assay measures multi-lineage reconstitution of hematopoiesis in irradiated transplant recipient mice. Thus this assay is routinely used to determine haematopoietic stem and progenitor cells (HSPCs) functionality in vivo. The principle of the method is to transplant bone marrow donor cells (derived from transgenic mice of choice) on C57BL6 background together with normal competitor bone marrow. In order to distinguish donor from competitor cells upon transplantation, usually competitor mice are congenic and carry the differential B cell antigen originally designated Ly5.1 and CD45.1.A typical competitive bone marrow transplantation experiment will contain two transplantation groups, donor (transgenic mice of choice and their controls) are transplanted in competition with normal competitors and engraftment efficiency is evaluated in both blood and bone marrow.
Keywords: Bone marrow transplantation, Hematopoesis, HSCs, Competitive, Stem cell engrafment
Materials and Reagents
Equipment
Software
Procedure
Part I: Bone marrow transplantation Before starting, it is important to understand that the following protocol is designed to test competitive bone marrow reconstitution of desired transgenic donors. Competition is always with wild-type congenic C57BL6 CD45.1 bone marrow cells. It is crucial to test the competition of appropriate wild-type control cells with the competitor and the comparison is always done between the tested donors and their wild-type controls.
Part II: Blood reconstitution analysis First analysis of total blood donor reconstitution is evaluated between 4-6 weeks following bone marrow transplantation. We usually start at 6 weeks and monitor reconstitution, once every two weeks up to 20 weeks.
Part III: Long-term bone marrow engraftment analysis
Recipes
Acknowledgments
The protocol was adapted from the previously published paper: Maryanovich et al. (2012). Our studies are supported in part by the Israel Science Foundation (ISF), USA-Israel Binational Science Foundation (BSF), German-Israel Foundation (GIF), Minerva Stiftung, MDM ICR Research Award, and Milgrom Award. A.G. is the incumbent of the Armour Family Career Development Chair of Cancer Research.
References
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