Fig. 1.

A, B Accumulated cell count and population doubling time (PDT) over five subcultures to understand the growth kinetics of SHED. C Senescence assay to assess the activity of betagalactosidase in SHED at SC3. D Trilineage differentiation of SHED at SC3. Adipogenesis was assessed via Red Oil staining and oil droplets formation was observed. Furthermore, chondrogenesis was evaluated using Alzarin Blue staining with the formation of proteoglycan networks. In addition osteogenesis was measured using Von Kossa staining with the presence of black precipitate. All micrographs were taken at 10x magnification and scale bar = 100μm. E Real-time PCR was performed to detect presence of representative genes related to adipogenesis, chondrogenesis and osteogenesis