The role of N-acetylcysteine supplementation on the oxidative stress levels, genotoxicity and lineage commitment potential of ex vivo murine haematopoietic stem/progenitor cells

Zariyantey Abd Hamid, Hui Y. Tan, Paik W. Chow, Khairul A.W. Harto, Chin Yi Chan, Jamaludin Mohamed

Research output: Contribution to journalArticle

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Abstract

Objectives: The ex vivo maintenance of haematopoietic stem/progenitor cells (HSPCs) is crucial to ensure a sufficient 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. This 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: This 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. The 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 significantly increased cell viability (P <0.050), while intracellular ROS levels significantly decreased at 0.25 and 0.5 µM (P <0.050). Moreover, DNA damage was significantly reduced at all NAC concentrations (P <0.050). Finally, the potential lineage commitment of the cells was not significantly affected by NAC supplementation (P >0.050). Conclusion: The findings of this study indicate that NAC supplementation may potentially overcome the therapeutic limitations of ex vivo-maintained HSPCs.

Original languageEnglish
Pages (from-to)e130-e136
JournalSultan Qaboos University Medical Journal
Volume18
Issue number2
DOIs
Publication statusPublished - 1 May 2018

Fingerprint

Acetylcysteine
Hematopoietic Stem Cells
Oxidative Stress
Malaysia
Reactive Oxygen Species
Cell Survival
Lymphoid Progenitor Cells
Therapeutic Human Experimentation
Comet Assay
Trypan Blue
Genomic Instability
Bone Marrow Cells
Maintenance
Staining and Labeling
Control Groups
Growth

Keywords

  • Cell lineage
  • DNA damage
  • Hematopoietic stem cells
  • N-acetylcysteine
  • Reactive oxygen species

ASJC Scopus subject areas

  • Medicine(all)

Cite this

The role of N-acetylcysteine supplementation on the oxidative stress levels, genotoxicity and lineage commitment potential of ex vivo murine haematopoietic stem/progenitor cells. / Abd Hamid, Zariyantey; Tan, Hui Y.; Chow, Paik W.; Harto, Khairul A.W.; Chan, Chin Yi; Mohamed, Jamaludin.

In: Sultan Qaboos University Medical Journal, Vol. 18, No. 2, 01.05.2018, p. e130-e136.

Research output: Contribution to journalArticle

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T1 - The role of N-acetylcysteine supplementation on the oxidative stress levels, genotoxicity and lineage commitment potential of ex vivo murine haematopoietic stem/progenitor cells

AU - Abd Hamid, Zariyantey

AU - Tan, Hui Y.

AU - Chow, Paik W.

AU - Harto, Khairul A.W.

AU - Chan, Chin Yi

AU - Mohamed, Jamaludin

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KW - Hematopoietic stem cells

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KW - Reactive oxygen species

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