The regulation of tendon stem cell distribution, morphology, and gene expression by the modulus of microfibers

Regulation of Tendon Stem Cell Behavior by Designed Nanoporous Topography of Microfibers

“…Polyamid-56 (PA56, Cathay Biotech Inc., China) was dissolved in hexafluoroisopropanol (HFIP, Sichuan Upper Fluorine Technology Co., LTD, China) solvent to produce 18, 20, 22 wt % solutions, respectively, as previously described. 22 The 3 PA56 solutions were then mixed with glycerol with the proportion of 6:1, 4:1, and 2:1 and stirred in a magnetic stirrer at room temperature at 300 rpm for 12 h to make the electrospinning solution. The crisscross fiber scaffold mats were fabricated by near-field electrospinning (Figure S1 and Table S1).…”

“…Rat Achilles tendon-derived stem cells were harvested and identified as TSCs. , TSCs at passage 4 to passage 7 with a density of 1.0 × 10 4 /mL were seeded on the surfaces of sterilized scaffolds (75 v/v % ethanol solution for 24 h, ultraviolet lamp for 6 h, complete culture medium for 12 h), fixed at the bottom of 24-well plate, and secured with rubber rings. The proliferation of TSCs was detected using the cell counting kit-8 (CCK 8) assay at the first, second, fourth, and seventh day.…”

“…For the immunofluorescence experiment, 22 TSCs were stained with primary antibodies collagen I (Col I, 1:500; ab34710) and collagen III (Col III, 1:1000; ab86829) from Abcam (Cambridge, UK), Nanog (1:200; sc-374103), OCT4 (1:200; Sc-5279) and (1:200; sc-21704) from (SANTA CRUZ, USA). The Cy3-labeled secondary antibody was incubated with cells at 37 °C for 1 h. The cell nuclei were reversely stained with DAPI (Bioindustrial, Israel) solution.…”