Comparison of Oxidative Stress Effects on Senescence Patterning of Human Adult and Perinatal Tissue-Derived Stem Cells in Short and Long-term Cultures

Abstract

Human Mesenchymal Stem Cells (hMSCs) undergo senescence in lifespan. In most clinical trials, hMSCs experience long-term expansion ex vivo to increase cell number prior to transplantation, which unfortunately leads to cell senescence, hampering post-transplant outcomes.Hydrogen peroxide (H2O2) in vitro represents a rapid, time and cost-effective tool, commonly used as oxidative stress tantalizing the stem cell ability to cope with a hostile environment, recapitulating the onset and progression of cellular senescence.Here, H2O2 at different concentrations (ranging from 50 to 400 μM) and time exposures (1 or 2 hours - h), was used for the first time to compare the behavior of human Adipose tissue-derived Stem Cells (hASCs) and human Wharton's Jelly-derived MSCs (hWJ-MSCs), as representative of adult and perinatal tissue-derived stem cells, respectively. We showed timely different responses of hASCs and hWJ-MSCs at low and high subculture passages, concerning the cell proliferation, the cell senescence-associated β-Galactosidase activity, the capability of these cells to undergo passages, the morphological changes and the gene expression of tumor protein p53 (TP53, alias p53) and cyclin dependent kinase inhibitor 1A (CDKN1A, alias p21) post H2O2 treatments.The comparison between the hASC and hWJ-MSC response to oxidative stress induced by H2O2 is a useful tool to assess the biological mechanisms at the basis of hMSC senescence, but it could also provide two models amenable to test in vitro putative anti-senescence modulators and develop anti-senescence strategies.

Effects of cryopreservation and long-term culture on biological characteristics and proteomic profiles of human umbilical cord-derived mesenchymal stem cells

“…De Witte et al reported that long-term expansion induced aging of hUC-MSCs exhibiting stable phenotype but reduced immunosuppressive properties from P4 to P12 [39]. Facchin et al reported that umbilical cord Wharton's Jelly-derived MSCs showed higher antioxidant ability to senescence than human adipose tissue-derived MSCs at high subculture passages, and they considered that the age of tissue donors is likely to be the main cause of senescence [40]. Moreover, recently, studies found that transcriptome and epigenetic regulations changes of hUC-MSCs occurred during long-term expansion [41,42].…”

Identification of senescent cells in multipotent mesenchymal stromal cell cultures: Current methods and future directions

“…Recent studies treating animal MSCs with ascorbic acid, or with an antioxidant cytokine C1q and tumor necrosis factor related protein 9 (CTRP9) [144], have also shown promise and need to be tested on human MSCs. In terms of selecting MSC tissue source for expansion, apart from tissue's propensity for certain lineage differentiation or trophic factor secretion, greater consideration should be given to MSC oxidative stress resistance [145] and desirably, longer onset of senescence. Even amongst perinatal cells, known to reach senescence later than adult tissue-MSCs [146], tissue specific differences exist, as shown recently in Kwon et al study [147].…”

Substance-P Restores Cellular Activity of ADSC Impaired by Oxidative Stress

“…Previous reports have induced oxidative stress in vitro by treating cells with H 2 O 2 for 24 h [ 25 , 26 ]. However, ADSCs appear to be particularly susceptible to H 2 O 2 -dependent oxidative stress, with 1 or 2 h of exposure being sufficient to impair ADSC activity [ 27 , 28 ]. Thus, in this study, we induced oxidative stress in ADSCs by exposing these cells to different concentrations of H 2 O 2 for 2 h ( Figure 1 A).…”

“…Moreover, a recent study corroborated the impairment of ADSCs by oxidative stress. Several studies have also indicated that, compared to other cells, stem cells are more susceptible to damage due to free radicals [ 27 ]. Thus, the effect of oxidative stress on stem cells should be taken into consideration for the application of stem cell therapy and endogenous regeneration.…”