30 min. The tissue was then transferred to a dissociation medium containing 3 U/ml collagenase F (Sigma-Aldrich) and 0.5 mg/ml BSA and incubated at 35°C for another 15 min to produce isolated ASM cells. Finally, the tissue was agitated with a fire-polished wide-bore glass pipette to release the cells. The isolated single ASM cells were used on the same day at room temperature (22–25°C).
