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

Natakarn Nimsanor; Jitrada Phetfong; Chotiros Plabplueng; Kulachart Jangpatarapongsa; Virapong Prachayasittikul; Aungkura Supokawej

doi:10.3892/etm.2017.5249

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

Exp Ther Med. 2017 Dec;14(6):5329-5338. doi: 10.3892/etm.2017.5249. Epub 2017 Oct 2.

Authors

Natakarn Nimsanor¹, Jitrada Phetfong², Chotiros Plabplueng², Kulachart Jangpatarapongsa², Virapong Prachayasittikul³, Aungkura Supokawej¹

Affiliations

¹ Department of Clinical Microscopy, Faculty of Medical Technology, Mahidol University, Salaya, Nakhon Pathom 73170, Thailand.
² Center for Research and Innovation, Faculty of Medical Technology, Mahidol University, Salaya, Nakhon Pathom 73170, Thailand.
³ Department of Clinical Microbiology and Applied Technology, Faculty of Medical Technology, Mahidol University, Salaya, Nakhon Pathom 73170, Thailand.

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.

Keywords: Wharton's jelly; cardiomyocytes; hydrogen peroxide; mesenchymal stem cells; oxidative stress.