Bone Marrow Transplantation in Mice to Study the Role of Hematopoietic Cells in Atherosclerosis

Abstract

Hematopoietic stem cell transplantation or bone marrow transplantation is a common approach to reconstitute the immune system of mice that have been subjected to marrow-ablative doses of radiation. This method can be used in the field of atherosclerosis to assess the contribution of hematopoietic cells of a desired genotype to disease pathogenesis. The engraftment of atherosclerosis-prone mice with donor cells that contain genetic alterations in cells of the innate or adaptive immune system has been invaluable to define the role of multiple gene products in atherosclerosis. Here, we describe the different steps involved in the bone marrow transplantation protocol along with specific guidelines regarding the theoretical and technical details of the procedure.

Viperin controls chikungunya virus–specific pathogenic T cell IFNγ Th1 stimulation in mice

“…For bone marrow chimera studies, 6-wk-old recipient mice were irradiated twice with 600 Rad (4 h apart) and injected (i.v.) with 4 × 10 6 donor bone marrow cells, as previously described (Sreeramkumar & Hidalgo, 2015). To test for successful adoptive bone marrow cell transfer, CD45.1 (WT) and CD45.2 ( Viperin −/− ) staining on 10 μl tail blood was assessed by flow cytometry 7.5 wk post engraftment (Fig S9), virus infection was performed by ventral injection of 1 × 10 6 pfu at 8 wk post engraftment.…”

Metformin Directly Suppresses Atherosclerosis In Normoglycemic Mice Via Haematopoietic Adenosine Monophosphate-Activated Protein Kinase (Ampk)

“…This approach is standard in the atherosclerosis literature and is usually interpreted as indicating myeloid specificity and even implied specificity for macrophages. [30][31][32][33][34] Recipient Ldlr -/mice were preconditioned with lethal c-irradiation, then rescued by adoptive transfer of bone marrow from mice deficient in AMPK (Prkab1 -/-) or littermate matched (Prkab1 þ/þ ) controls. BMT was performed at age 12 weeks, and metformin treatment from age 14 weeks to age 25 weeks, allowing a prolonged period on metformin after BMT, when there was a cull for analysis as for the first cohort (Figure 2A).…”

CETP Expression in Bone-Marrow-Derived Cells Reduces the Inflammatory Features of Atherosclerosis in Hypercholesterolemic Mice

“…To investigate the impact of macrophages in atherosclerosis, bone marrow transplantation in mice offers a powerful and effective strategy. This approach allows for the generation of chimeric animals with a hematopoietic compartment carrying the donor's genetic background, which enables the identification of the role of the genetic profile of these cells in the development of atherosclerosis [10]. Consistent evidence has shown that the reconstitution of Apoe −/− mice with wild-type bone marrow cells results in protection from high lipid levels and atherosclerosis, and, conversely, wild-type mice reconstitution with Apoe −/− bone marrow cells exacerbates atherosclerosis [11,12].…”