Bashir B, Choudhery MS, Hussain I. Hypoxic preconditioning restores the regenerative potential of aged adipose tissue derived mesenchymal stem cells. World J Stem Cells 2026; 18(5): 117241 [DOI: 10.4252/wjsc.v18.i5.117241]
Corresponding Author of This Article
Mahmood S Choudhery, PhD, Associate Professor, Department of Human Genetics and Molecular Biology, University of Health Sciences, Khayaban-e-Jamia, Lahore 54000, Punjab, Pakistan. ms20031@yahoo.com
Research Domain of This Article
Cell Biology
Article-Type of This Article
research-article
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World J Stem Cells. May 26, 2026; 18(5): 117241 Published online May 26, 2026. doi: 10.4252/wjsc.v18.i5.117241
Hypoxic preconditioning restores the regenerative potential of aged adipose tissue derived mesenchymal stem cells
Bushra Bashir, Mahmood S Choudhery, Ijaz Hussain
Bushra Bashir, Ijaz Hussain, Department of Dermatology, King Edward Medical University, Lahore 54000, Punjab, Pakistan
Mahmood S Choudhery, Tissue Engineering and Regenerative Medicine Laboratory, King Edward Medical University, Lahore 54000, Pakistan
Mahmood S Choudhery, Department of Human Genetics and Molecular Biology, University of Health Sciences, Lahore 54000, Punjab, Pakistan
Author contributions: Choudhery MS designed the study; Basir B performed the experimental procedures and collected data; Bashir B and Hussain I performed the data analysis and interpretation; Choudhery MS and Hussain I supervised the study, provided critical input throughout the research process, and reviewed the manuscript. All authors participated in manuscript writing, revision, and approved the final version of the manuscript for submission.
Institutional review board statement: The study was conducted in accordance with ethical standards and approved by the Advanced Studies & Research Board of King Edward Medical University (No. 2744/KEMU/2016). All procedures involving human samples complied with applicable guidelines and regulations, and informed consent was obtained from all donors prior to sample collection.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: The data supporting the findings of this study has been included in the study.
Corresponding author: Mahmood S Choudhery, PhD, Associate Professor, Department of Human Genetics and Molecular Biology, University of Health Sciences, Khayaban-e-Jamia, Lahore 54000, Punjab, Pakistan. ms20031@yahoo.com
Received: December 2, 2025 Revised: January 6, 2026 Accepted: March 12, 2026 Published online: May 26, 2026 Processing time: 174 Days and 14.4 Hours
Abstract
BACKGROUND
Mesenchymal stem cells (MSCs) hold great promise for aesthetic dermatology due to their self-renewal, multipotency, and paracrine effects. However, MSCs derived from aged donors exhibit limited therapeutic efficacy in autologous applications due to their impaired regenerative potential. Hypoxic preconditioning has been explored as an effective strategy to improve the potential of stem cells. In the current study, we aim to prime cultured aged adipose tissue derived MSCs (ADMSCs) under hypoxia to augment their regenerative potential.
AIM
To evaluate the effect of hypoxic preconditioning on enhancing the regenerative capacity of aged ADMSCs.
METHODS
ADMSCs were isolated from 12 young (≤ 20 years) and 12 aged (≥ 60 years) donors. ADMSCs isolated from both donor groups were cultured under normoxic conditions (21% O2), while aged ADMSCs were additionally subjected to hypoxic preconditioning at 3% O2 for 48 hours at each passage. We performed various assays to determine the effect of hypoxia on growth kinetics; cumulative population doublings, doubling time, clonogenic potential (colony forming unit assay, plating efficiency), differentiation (osteogenic and adipogenic), migration (in vitro scratch assay), pro-angiogenic (tube formation assay), senescence (senescence-associated β-galactosidase staining), and expression of various genes using reverse transcriptase quantitative polymerase reaction.
RESULTS
Hypoxic preconditioned aged ADMSCs exhibited higher cumulative population doublings, lower doubling time, decreased senescence, enhanced in vitro wound closure, improved colony formation, greater angiogenic, osteogenic, and adipogenic differentiation potential compared with aged-normoxic ADMSCs. Gene expression analysis showed upregulation of protein kinase B, sirtuin 1, insulin-like growth factor 1, vascular endothelial growth factor and stromal cell-derived factor-1 genes, concurrently with downregulation of BAX, BAK1, P53, P21 and P16 genes.
CONCLUSION
Hypoxic preconditioning enhances regenerative potential of aged ADMSCs. This strategy may be promising for autologous regenerative medicine applications, particularly for autologous therapy.
Core Tip: Mesenchymal stem cells (MSCs) exhibit age-related decline in their regenerative potential. Hypoxic preconditioning has been found to be an effective strategy to augment their regenerative potential. Aged adipose tissue derived MSCs subjected to hypoxic preconditioning (3% O2) for 48 hours exhibited enhanced proliferative potential, clonogenicity, differentiation capability, concurrent with reduced senescence. These findings suggest that hypoxic-preconditioning is a promising therapeutic option to enhance the regenerative potential of aged adipose tissue derived MSCs for autologous use.
