The invention discloses a method for culturing adult mouse cardiac fibroblasts. The method comprises the following steps: taking SPF grade 8-12 weeks-old mice, and sterilizing the material after cervical dislocation; taking out the heart of the mice and placing the mice in a culture dish containing pre-cooled calcium-free HBSS, extruding the residual blood in the heart, removing the heart atrium,left and right auricula, and pericardium, preserving the ventricle, transferring the material to a new culture dish containing the pre-cooled calcium-free HBSS, cutting the heart, transferring the material to a centrifuge tube, and after the cardiac tissue is precipitated, sucking the HBSS by a pipette; adding a collagenase digestion solution in the centrifuge tube for digestion, and then resuspending the material to obtain a cell suspension; spreading the material into a first culture dish and performing standing culture; resuspending the cells, discarding a supernatant by centrifugation, spreading the resuspended cells in the medium to a second culture dish, and adding the original first culture dish to the new medium. The culture method is simple and easy, has good stability, high repeatability and uniform morphology, and the obtained fibroblasts grow well.