Research Paper Volume 16, Issue 5 pp 4904—4919

Figure 1. Late passaged UCMSCs showed senescence-associated changes and impaired biological functions. (A) Population double time (PDT) of UCMSCs during long-term expansion. (B) The cell size of early and late passaged UCMSCs was measured using forward scatter (FSC) and side scatter (SSC) by flow cytometry. (C) Immunophenotypes characterization of early and late passaged UCMSCs. (D) SA-β-gal staining of early and late passaged UCMSCs. (E) Quantification of the SA-β-gal staining positive cells (n = 3). (F) Western blot analysis of p16 expression in UCMSCs of passages 4, 8, and 11. (G) Early and late passaged UCMSCs were induced to osteogenic (Ost) or adipogenic (Ado) differentiation. Osteogenic efficiency was evaluated by alkaline phosphatase (ALP) staining; adipogenic efficiency was evaluated by oil red O staining. (H) The differentiation rate was quantized. (I) Wound healing assay was performed for early and late passaged UCMSCs and (J) migrated cell numbers were quantified in 0 hours, 6 hours, 12 hours and 24 hours (n = 3). ^***p < 0.001.