Role of Glutathione in Balancing Total Antioxidant Status and Generation of Reactive Oxygen Species During Hepatic Differentiation of Bone-Marrow-Derived Mesenchymal Stem Cells



Allameh A¹ ; Ahmadiashtiani HR¹ ; Rezapour M¹ ; Rastegar H²
Source: Journal of Stem Cells Published:2018

Abstract

Differentiation of mesenchyal stem cells (MSCs) into biologically active hepatocytes depends on several factors. Glutathione (GSH) and antioxidant enzymes play major role in maintaining the balance between oxidative damage factors and antioxidants during the differentiation. Therefore by adding GSH modifiers to the culture media, we examined the impact of GSH on the process of MSC differentiation into hepatocyte-like cells. In the present study differentiation of MSCs into hepatocyte-like cells was induced using a 2-step protocol. Then production of reactive oxygen species (ROS), cellular GSH, GSH peroxidase (GPX), GSH reductase (GR) and total antioxidant capacity (TAC) were determined on days 2, 7 and 14 of differentiation onset. Besides, the experiment was carried out in cells treated with glutathione modifiers namely; buthionine sulfoximine (BSO) or N-acetylcysteine (NAC). Depletion in GSH and GPX due to BSO treatments resulted in elevation of ROS production. In contrast, ROS was substantially decreased in cells having high levels of GSH and GPX following NAC treatments. The results showed that during differentiation, the cell proliferation was affected by ROS alterations. Cell proliferation was enhanced when treated with NAC which also increased GSH, GPX and TAC. As a consequence of GSH elevation by NAC, there was a substantial increase in antioxidant activity of the cells undergoing differentiation. This data show that ROS generation and TAC status can be balanced during the process of hepatic differentiation of MSCs by inducing GSH biosynthesis using NAC in the culture media. This action is associated with improved cell proliferation and differentiation. © 2018 Nova Science Publishers, Inc.