Oxidative status predicts quality in human mesenchymal stem cells

Abstract

BackgroundHuman bone marrow-derived mesenchymal stem cells (MSC) are adult progenitor cells with great potential for application in cell-based therapies. From a cell-based therapy perspective, there are two limitations to MSC use: (1) these therapies require large numbers of cells, and long-term expansion of MSC in vitro promotes replicative senescence; and (2) patient variability is a challenge for defining MSC quality standards for transplantation. This study aimed to determine whether low or high oxidative status of MSC correlate with changes in cell expansion and differentiation potentials.MethodsWe investigated functional aspects of mitochondria, such as cell metabolic activity indicators and expression of antioxidant enzymes. Furthermore, we tested if senescence-induced changes in oxidative status of MSC could be counteracted by methylene blue (MB), an alternative mitochondrial electron transfer known to enhance cell bioenergetics.ResultsMSC isolated from donors of the same age showed distinctive behavior in culture and were grouped as weak (low colony-forming units (CFU) and a short life in vitro) and vigorous MSC (high CFU and a long life in vitro). In comparison to weak MSC, vigorous MSC had oxidative status characterized by lower mitochondrial membrane potential, lower mitochondrial activity, and fewer reactive oxygen species production, as well as reduced mitochondrial biogenesis. Vigorous MSC had a significantly higher expansion potential compared to weak MSC, while no differences were observed during differentiation. MB treatment significantly improved expansion and differentiation potential, however only in vigorous MSC.ConclusionsTogether, these results demonstrate the importance of mitochondrial function in MSC in vitro, and that cells with low oxidative status levels are better candidates for cell-based therapies.

Mitochondrial Oxidative Stress Reduces the Immunopotency of Mesenchymal Stromal Cells in Adults With Coronary Artery Disease

“…50 Further we show that atherosclerotic-MSCs have higher levels of total intracellular ROS (a negative predictor of the MSCs expansion potential 51 ) and, in particular, elevated mitochondrial ROS levels that correlate with their reduced in vitro immunopotency. Considering that MSCs may play a role in inhibiting inflammation and stabilizing vulnerable atherosclerotic plaques, 52 and the implications of mitochondria to atherosclerosis development, 53 our findings provide the rationale for testing whether restoring mitochondrial dysfunction in atherosclerotic-MSCs could impact atherosclerosis progression reducing the risk of acute events.…”

Cryopreserved H2 O2 -preconditioned human adipose-derived stem cells exhibit fast post-thaw recovery and enhanced bioactivity against oxidative stress

“…Finally, it has recently been described that a preserved low oxidative status reduces oxidative stress in cells, maintaining a prolonged resourceful homeostasis in MSCs, indicating that oxidative status could be a predictor of expansion potential in MSCs (Bertolo et al, ). From this perspective, we have found that our HC016 cells showed lower basal mitochondrial activity than their corresponding control ASCs.…”

Rapid Detection of Senescent Mesenchymal Stromal Cells by a Fluorescent Probe

“…Interestingly, the targeted elimination of senescent cells using senolytics was shown to greatly improve the proliferation of cells . In order to achieve this, it will be of utmost importance to monitor the development of senescence . Thus, identification of the early onset of senescence is key to achieving the targeted elimination of senescent cells, which would greatly improve the quality of MSCs for therapeutic purposes .…”