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.
