A method for preparing an agonist for improving boar sperm motility includes: (1) collecting and infiltrating a testicular tissue of a young boar 3-5 days after born; (2) washing the tissue with PBS; (3) incubating the tissue in a cell culture medium, and centrifuging the cell suspension to remove a supernatant; (4) adding hyaluronidase and collagenase IV followed by shaking, and adding the cell culture medium and centrifuging to remove a supernatant; (5) adding trypsin and deoxyribonuclease followed by shaking, and adding the cell culture medium; (6) filtering the cell suspension by a cell sieve; (7) centrifuging to remove a supernatant, and adding the cell culture medium; (8) culturing and passaging the cells; (9) culturing and storing the cell suspension. The invention has the advantages of good application effect, low cost and simple production process.