〔Abstract〕 To establish an in vitro culture model ofrat corpora cavernous smooth muscle cells (CCSMCs) using a modified tissue block adherence method. Methods Corpus cavernosum smooth muscle tissue was digested with collagenase type I and subsequently cultured using an adherent method. Cells were purified via differential adhesion and identified through immunofluorescence and Western blotting. Results CCSMCs began to emerge from the tissue block after 3 days, increased significantly by day 7, and converged by day 12. Post-passage, CCSMCs exhibited strong proliferation and a “peak-to-valley” phenomenon. After purification, the cells tested positive for α-smooth muscle actin (α-SMA), confirming the successful establishment of the in vitro culture model. Conclusion The modified tissue block adherence method is a cost-effective and efficient way to obtain high-purity CCSMCs.