Inhibitory effect of oxidative damage on cardiomyocyte differentiation from Wharton's jelly‑derived mesenchymal stem cells

Abstract

Ischemic heart diseases are a serious health problem worldwide. The transplantation of mesenchymal stem cells (MSCs) has been investigated in numerous clinical trials on various other diseases due to the self-renewal capacity of these cells and their potential to differentiate into a variety of cell types. The presence of excess reactive oxygen species in injured myocardium causes cardiac dysfunction and leads to inefficient repair of the heart. The poor outcomes of stem cell transplantation have been suggested to result from residual oxidative damage affecting the transplanted cells. The aim of the present study was to compare the effects of hydrogen peroxide (H2O2) on Wharton's jelly-derived MSCs (WJ-MSCs) and bone marrow-derived MSCs (BM-MSCs) in vitro, in order to provide information useful for the future selection of MSC types for cardiac differentiation and transplantation. H2O2 at concentrations of 200, 500 and 1,000 µM was applied to WJ-MSCs and BM-MSCs under cardiogenic differentiation conditions. The morphology of MSCs treated with H2O2 was similar to that of untreated cells, whereas the cell density decreased in direct association with the dose of H2O2. Cardiac differentiation markers were then evaluated by immunofluorescence analysis of GATA4 and cardiac troponin T (cTnT). The fluorescence intensity levels of the two markers were identified to be diminished by increasing doses of H2O2 from 500 to 1,000 µM. The expression levels of homeobox protein Nkx2.5, cTnT and cardiac α-actin were also examined, and were identified to be low in the WJ-MSCs treated with 1,000 µM H2O2, which was similar to the findings observed in BM-MSCs. These results suggested that oxidative stress affects cardiomyocyte differentiation via the downregulation of cardiac genes and cardiac proteins. Furthermore, it should be noted that there was a marked difference in the effect depending on the source of MSCs. This evidence provided supportive information for the use of stem cells in transplantation.

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

“…Here, we found that in both cell populations, H 2 O 2 not only inhibited cell proliferation in a dose-dependent fashion, but it also affected the persistence of the anti-proliferative effect after the end of exposure (up to 48 or 72 h) in a manner that was dependent upon the duration (1 or 2 h) of the treatment itself. These results are coherent with the reduction of cell vitality observed in hWJ-MSCs 43 - 45 and hASCs 39 , 42 treated with higher concentrations of H 2 O 2 (until 2000 μM) but at low subcultured passages. We also observed a growth slowdown of cells treated with H 2 O 2 400 μM, and a cell inability to re-plate.…”

“…Moreover, H 2 O 2 treatment provokes DNA breaks 41 , raises SA β-Gal positive cells 42 , alters the expression of senescent marker genes, as well as p53 , p21 , mitogen-activated protein kinase 14 ( MAPK14 , alias p38 ) and sirtuin 1 ( SIRT1 ) 38 , 39 , 42 , and increases apoptosis with a decline of pro-survival gene expression 38 . It has been recently shown that also human Wharton's Jelly-derived MSCs (hWJ-MSCs) treated with H 2 O 2 undergo premature senescence at early culture passages: these cells show typical changes in morphology 43 , slow their proliferation 44 , 45 , result positive for SA β-Gal 43 , 44 , express typical senescence 43 and pro-apoptotic gene markers, while displaying a down-regulation of survival genes 44 , 46 .…”

Overexpression of anti-fibrotic factors ameliorates anti-fibrotic properties of Wharton's jelly derived mesenchymal stem cells under oxidative damage

“…Umbilical cords were collected from mothers with normal labor as previously described (28). All participants have read and signed the informed consent, which was approved by the Mahidol University Institutional Review Board (protocol no.…”

“…WJ-MSCs in all experiments were performed as previously described (28). WJ-MSCs were characterized according to the minimal criteria for defining MSCs, stated by the International Society for Cellular Therapy (29).…”

“…For mesodermal differentiation properties of MSCs, WJ-MSCs were cultured with osteogenic and adipogenic differentiation medium (Stem Cell Technologies, Vancouver, Canada) as previously described (28). WJ-MSCs were cultured with differentiation medium for 35 days.…”