Fig. 2

Isolation of GSCs in PL10% + GH medium and phenotypic properties. A Microscopic views of gingival fibroblasts (GFs) and GSCs isolated at 1 week and 3 weeks of explants and enzymatic digestion. GSCs retain their fibroblastic morphology in both FCS and PL media, but PL-isolated cells are larger. B CD29, actin and vimentin immunostaining of S-GSCs and L-GSCs. Cells were grown for 72 h on coverslips coated with gelatin 0.2%. C FACS analysis of MSC surface markers showed that both S-GSCs and L-GSCs were positive for CD29, CD44, CD90 and CD105. D DNA lesion detection was very low using Anti-H2AX staining in S-GSCs and L-GSCs. E and F Colony-forming unit-fibroblast (CFU-F) assay: Crystal violet staining showed that L-GSCs produce thicker and more robust colonies than S-GSCs, with a nearly similar number. (G and H) RT-qPCR gene expression analysis of GH-R1, GH-RU and TGFβ-R in both S-GSCs and L-GSCs. Gene expression was confirmed by migration of amplified DNA on an agarose gel, and the relative expression of the 3 genes was determined by normalisation with the SDHA and UBC as reference genes. No significant difference was found between S-GSCs and L-GSCs. Scale bars: A 1000 µM, 400 µM, B 400 µM, D 20 µM