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. 2020 Dec 15;3:774. doi: 10.1038/s42003-020-01514-y

Fig. 1. In vitro culture expansion of human mesenchymal stem cells (hMSCs) results in cellular senescence and breakdown of cellular homeostasis.

Fig. 1

a Alteration of hMSC morphology during culture expansion. b SA-β-gal activity and c SA-β-gal staining in hMSCs at early passage and late passage. d Comet assay demonstrates DNA damage of hMSCs during culture expansion. e Population doubling (PD) time increased for long-term cultured hMSCs. f Colony-forming unit-fibroblast (CFU-F) ability decreased during culture expansion of hMSCs. g mRNA levels of stem cell genes and h mRNA levels of cell cycle genes in hMSCs at late passage compared to cells at early passage. i Cell cycle analysis of hMSCs via flow cytometry. j Autophagic gene expression of hMSCs during culture expansion. k Basal autophagic flux was reduced in hMSCs at late passage compared to early passage. Late passage of hMSCs: passage 12 (P12), early passage of hMSCs: passage 5 (P5). Biological replicates (n): n = 3 for the tests. Scale bar: 100 µm. *p < 0.05; **p < 0.01; ***p < 0.001.