The single-anastomosis heart-lung transplantation method in the rat developed by Lee and associates has been examined and has proven to be simple and reproducible. Technical procedures are discussed emphasizing those procedures that require special caution to ensure successful graft function. The donor and recipient rat strain combination has been found to influence the success of the transplantation procedure. Rat strain combinations used in our laboratories resulted in different percentages of donor heart survival. Using Lewis rat (RT1 1/1) recipients, 68% of Brown Norway rat (RT1n/n) donor hearts and 97% of NBR rat (RT1 1/1) donor hearts survived. However, only 29% of Fischer rat (RT1 1/1) donor hearts survived longer than 2 days when transplanted into Lewis rat recipients. In our laboratories, the Lee method of single-anastomosis heart-lung transplantation has been used successfully to evaluate whether immunoregulatory compounds alter heart allograft rejection.