Primary cultures (passages 4∼6) wer

Primary cultures (passages 4∼6) were grown as three- dimensional fibroblast-populated collagen lattices (FPCLs). Specifically, collagen lattices were prepared by mixing cells with a neutralized solution of collagen type I (eight parts collagen type I, 2.9 mg/ml [Sigma-Aldrich, St. Louis, MO, USA] one part 10× DMEM), so that the final collagen and cell concentrations were 2.0 mg/ml and 1×105 cells/ml, respectively. The cell-collagen mixture was then aliquoted into 24-well culture dishes (0.5 ml/well). Once polymerized (1 h, 37oC), DMEM 10% FBS was added on top of the FPCLs in each well. After incubation for 2 days, the attached FPCLs were mechanically released from the sides of the culture plates. To induce FPCL contraction, six sites on the FPCL were then treated with bipolar RF (1 MHz, 23 W) (Fig. 2A, B). Digital images of the contracting FPCL were captured at various time points (24 h, 48 h) during the next 2 days (Fig. 2C). Contraction of lattices was determined by averaging the longest and shortest diameter of each lattice measured with calipers. The diameters were calculated and represented as the mean and standard deviation of triplicate lattices (Fig. 2D, E).

Primary cultures (passages 4~6) were grown as three- dimensional fibroblast-populated collagen lattices (FPCLs). Specifically, collagen lattices were prepared by mixing cells with a neutralized solution of collagen type I (eight parts collagen type I, 2.9 mg / ml [Sigma-Aldrich, St. Louis, MO, USA] + one part 10 × DMEM), so. that the final collagen and cell concentrations were 2.0 mg / ml and 1 × 105 cells / ml, respectively. The cell-collagen mixture was then aliquoted into 24-well culture dishes (0.5 ml / well). Once polymerized (1 h, 37oC), DMEM + 10% FBS was added on top of the FPCLs in each well. After incubation for 2 days, the attached FPCLs were mechanically released from the sides of the culture plates. To induce FPCL contraction, six sites on the FPCL were then treated with bipolar RF (1 MHz, 23 W) (Fig. 2A, B). Digital images of the contracting FPCL were captured at various time points (24 h, 48 h) during the next 2 days (Fig. 2C). Contraction of lattices was determined by averaging the longest and shortest diameter of each lattice measured with calipers. The diameters were calculated and represented as the mean and standard deviation of triplicate lattices (Fig. 2D, E).

Primary cultures (passages 4 ∼ 6) were grown as three - dimensional fibroblast-populated collagen lattices (FPCLs). Specifically,, Collagen lattices were prepared by mixing cells with a neutralized solution of collagen type I (eight parts collagen type. I 2.9, mg / ml [Sigma-Aldrich St. Louis,,, MO USA] one part 10 × DMEM), so that the final collagen and cell concentrations. Were 2.0 mg / ml and 1 × 105 cells / ml respectively. The, cell-collagen mixture was then aliquoted into 24-well culture dishes (0.5. Ml / well). Once polymerized (1, H 37oC), DMEM 10% FBS was added on top of the FPCLs in each well. After incubation for, 2 days. The attached FPCLs were mechanically released from the sides of the culture plates. To induce, FPCL contractionSix sites on the FPCL were then treated with bipolar RF (1, MHz 23 W) (Fig. 2A B). Digital, images of the contracting. FPCL were captured at various time points (24 h 48 h), during the next 2 days (Fig. 2C). Contraction of lattices was determined. By averaging the longest and shortest diameter of each lattice measured with calipers.The diameters were calculated and represented as the mean and standard deviation of triplicate lattices (Fig,. 2D E).