Optimal good manufacturing practice-compliant production of dental follicle stem cell sheet and its application in Sprague-Dawley rat periodontitis

Figure 3 The effect of inoculation cell concentration on the characteristics of cell sheet. A: Acridine orange/propidium iodide staining and quantitative analysis of dead cells in cell sheets at varying initial cell concentrations. Scale bar = 100 μm; B: Final cell count in cell sheets at various seeding densities; C: Quantitative reverse-transcription polymerase chain reaction analysis of osteogenic, chondrogenic, odontogenic, and senescence-related genes in cell sheets with different seeding densities; D: Quantitative assessment of β-galactosidase staining in cell sheets at varying initial cell concentrations; E: Relative telomere length in cell sheets cultured at various cell concentrations; F: Measurement of hydroxyproline content in cell sheets at different initial cell concentrations; G: Measurement of glycosaminoglycan content in cell sheets at different initial cell concentrations; H: Alizarin red staining of cell sheets cultured at different initial cell densities. Scale bar = 100 μm. Data are presented as mean ± SD, n = 3. ^aP < 0.05; ^bP < 0.01; ^cP < 0.001. ALP: Alkaline phosphatase; RUNX-2: Runt-related transcription factor 2; CAP: Cartilage associated protein; COL-1: Type I collagen; OPN: Osteopontin; OCN: Osteocalcin; β-GAL: β-galactosidase; HYP: Hydroxyproline; sGAG: Sulfated glycosaminoglycan.