Hbsmcs

Following the method described previously by Park et al. [27 ], hBSMCs (catalog no. 4310; ScienCell, Carlsbad, CA, USA) and ICCs (FDCC, Shanghai, CHN) were seeded at 8×10⁴ cells/well in 6-well silicone elastomer-bottomed culture plates coated with type I collagen (Bioflex; Flexcell, Hillsborough, NC) and grown to 80% confluence in DMEM/10% FBS and a 5% CO₂ humidified atmosphere at 37°C. Cells of hBSMCs and hBSMCs/ICCs of co-culture were then subjected to continuous cycles of stretch-relaxation using a computer-driven, stretch-inducing device (Strain UnitFX-5000, Flexcell). Every cycle consisted of 5 s of stretch and 5 s of relaxation (0.1 Hz), and mechanical stretch was conducted under conditions of 5%, 10%, 15%, and 20% elongation (0.1 Hz) for 0, 2, 6, 12, and 24 h. Our results showed that mechanical stretch under conditions of 10% elongation (0.1 Hz) for 6 h is the best, so all subsequent experiments were carried out under those conditions. Imatinib (Monmouth Junction, NJ, USA) was dissolved in 0.01% dimethylsulfoxide. The treated concentration of imatinib was 10 μM, based on the method described by Kubota et al. [28 (link)].