Col 1 antibody

The hDFs were cultured at 37 °C, 5% CO₂ in high-glucose DMEM containing 1% antibiotic antimycotic and 10% FBS. The hDFs were seeded in a 24-well culture plate at a density of 1 × 10⁴ cells/well. The multifunctional PDO threads (40 mm) were treated on 24-well inserts in each well for 24 h. The live/dead fluorescence images and cell viability were measured using the Calcein AM/EthD-1 staining kit and the CCK-8 assay according to the provided instructions, respectively.
The hDFs were seeded at a density of 3 × 10⁴ cells on 24-well plates and the PDO threads were processed and incubated for 24 h. After processing, the cells were rinsed with PBS solution and fixed with 4% PFA solution for 15 min, permeabilized with 0.3% triton for 30 min, and finally blocked by 1% BSA solution for 30 min. Cells were probed with a COL1 antibody (Santa Cruz Biotechnology; Santa Cruz, CA, USA) in 5% BSA/0.1% Triton X-100 solution at 4 °C overnight. After rinsing with PBS solution and treated secondary antibody (Alexa-Fluor 568 goat anti-rabbit; Molecular Probes Inc., Eugene, OR, USA) in 5% BSA/0.1% Triton X-100 for 1 h at room temperature. Following this, they were rinsed 3 times with the PBS solution and mounted using Vectashield mounting medium/DAPI (Vector, CA, USA). The cells were observed using fluorescent microscopy.