The Role of N-Acetylcysteine Supplementation on the Oxidative Stress Levels, Genotoxicity and Lineage Commitment Potential of Ex Vivo Murine Haematopoietic Stem/Progenitor Cells
Journal Title: Sultan Qaboos University Medical Journal - Year 2018, Vol 18, Issue 2
Abstract
Objectives: Te ex vivo maintenance of haematopoietic stem/progenitor cells (HSPCs) is crucial to ensure a sufcient supply of functional cells for research or therapeutic applications. However, when exposed to reactive oxygen species (ROS) in a normoxic microenvironment, HSPCs exhibit genomic instability which may diminish their quantity and quality. Tis study aimed to investigate the role of N-acetylcysteine (NAC) supplementation on the oxidative stress levels, genotoxicity and lineage commitment potential of murine haematopoietic stem/progenitor cells (HSPCs). Methods: Tis study was carried out at the Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia, between June 2016 and July 2017. Bone marrow cells were isolated from nine mice and cultured in a growth medium. Various concentrations of NAC between 0.125–2 µM were added to the culture for 48 hours; these cells were then compared to non-supplemented cells harvested from the remaining three mice as the control group. A trypan blue exclusion test was performed to determine cell viability, while intracellular ROS levels and genotoxicity were determined by hydroethidine staining and comet assay, respectively. Te lineage commitment potential of erythroid, myeloid and pre-B-lymphoid progenitor cells was evaluated via colony-forming cell assay. Results: NAC supplementation at 0.25, 0.5 and 2 µM signifcantly increased cell viability (P <0.050), while intracellular ROS levels signifcantly decreased at 0.25 and 0.5 µM (P <0.050). Moreover, DNA damage was signifcantly reduced at all NAC concentrations (P <0.050). Finally, the potential lineage commitment of the cells was not signifcantly affected by NAC supplementation (P >0.050). Conclusion: Te fndings of this study indicate that NAC supplementation may potentially overcome the therapeutic limitations of ex vivo-maintained HSPCs.
Authors and Affiliations
Zariyantey A. Hamid, Hui Y. Tan, Paik W. Chow, Khairul A. W. Harto, Chin Y. Chan, Jamaludin Mohamed
Keywords
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