Secretome of Human Fetal Mesenchymal Stem Cell Ameliorates Replicative Senescen

Autologous mesenchymal stem cells (MSC) are widely used cell source for cell-based tissue repair and regeneration, but replicative senescence and the associated loss of cellular activity during in vitro expansion limit their therapeutic potential. How to preserve or even enhance the proliferation and differentiation ability of MSC from aged donors without genetic modification remains a challenge to meet clinical need. MSC isolated from human fetal tissues (hFMSC) exhibit higher proliferation and differentiation activities even in prolonged in vitro culture, which might be modulated by autocrine/paracrine action. In the present study, we hypothesized that the bioactive factors secreted by hFMSC, collectively named as hFMSC secretome (HFS), could possess beneficial effect on human adult MSC (hAMSC) undergoing replicative senescence, thus promoting their capability of proliferation and differentiation. HFS was prepared by centrifugation of hFMSC conditioned medium, followed by column-based concentration, and the total protein content of the HFS was quantified to standardize treatment concentration. When compared with hAMSC secretome (HAS), HFS treatment significantly reduced senescence associated-β-galactosidase expression and activity (senescence marker) and enhanced cell proliferation and osteogenic differentiation potential of hAMSC in prolonged in vitro culture. Cellular studies revealed concomitant activation of sirt1 and foxo3a in hAMSC after HFS treatment, which was associated with upregulation of p21 and downregulation of bax and p53. The changes of these senescence associated markers suggested that HFS, but not HAS, could ameliorate replicative senescence of hAMSC in vitro. In nude mice, HFS pretreatment restored the osteogenic ability of senescent hAMSC. Tumor xenograft model revealed that HFS did not promote tumor growth. In conclusion, this study suggests that HFS could be an effective and safe method to overcome replicative senescence and facilitate the therapeutic potential of hAMSC.