Techniques for perfusion and storage of heterotopic heart transplants in mice

Abstract

Organ pretreatment prior to transplantation has assumed increasing importance. We studied the ability to preserve and perfuse hearts in well-defined genetic mouse models, prior to heterotopic engraftment. Hearts were manually perfused through the aortic root and stored in a variety of cold solutions or were perfused using a continuous perfusion pump. Different electrolyte solutions, mouse strains, perfusion, and storage times as well as perfusion volumes were studied. Treated hearts were than transplanted heterotopically and short- and long-term function assessed. Hearts stored for more than 1 hour in cold solutions (saline 0.9%, or lactated Ringer's) failed to function. Hearts perfused with 0.25-0.4 ml of cold solution for a maximum of 30 minutes functioned well after transplantation. C3H (H-2k) mice provided the most resilient hearts. We conclude that short-term perfusion or storage of mouse hearts is feasible and should provide an excellent model for the study of organ pretreatment with monoclonal antibodies or other agents prior to transplantation.

Aquaporin 4 blockade improves survival of murine heart allografts subjected to prolonged cold ischemia

“…This was not due to the systemic lymphocyte depletion as the numbers of spleen T lymphocytes were comparable in control and AER-271 injected mice on day 3 and at the end of the treatment (16.5 ± 0.8 × 10 6 vs 14.0 ± 1.0 × 10 6 for CD4 T cells and 7.3 ± 0.3 × 10 6 vs 7.8 ± 0.5 × 10 6 for CD8 T cells). Similar improved graft survival and decreased priming of donor-reactive T cells were observed for heart allografts subjected to 8 h CIS in Ringer’s lactate, a solution commonly used in rodent organ transplantation (Figure 2C–D) (24, 25). Despite impaired T cell alloimmunity, recipients treated with AER-271 developed donor-reactive IgG alloantibodies (Figure 2E).…”

Differential depletion of total T cells and regulatory T cells and prolonged allotransplant survival in CD3Ɛ humanized mice treated with polyclonal anti human thymocyte globulin

“…Graft function was evaluated daily accordingly to the heart beat score evaluation as reported in [20], scoring system: 0: no organ function, 1: fibrillation, only visible through magnification, 2: poor or partial organ function, 3: impairment in frequency or intensity of heart beating, 4: physiological organ function.…”

Early detection of rejection and assessment of cyclosporine therapy by 111In antimyosin imaging in mouse heart allografts.

“…Prolonged storage of donor hearts in a cold solution has been shown to cause mechanical dysfunction. 25 The results of our acute experiment using antimyosin clearly indicated that storage of donor hearts in a cold solution causes significant ischemic myocyte necrosis. Grafts with an isehemic time of more than 90 minutes showed significantly increased antimyosin uptake in comparison with grafts with an ischemia time of less than 60 minutes.…”