Alpha Lipoic Acid Promotes the Proliferation, Motility, and Antioxidant Defense System of Human Adipose-Derived Stromal/Stem Cells

Abstract

Adipose-derived stromal/stem cells (ASCs) are a readily accessible mesenchymal stem cell population with high regenerative and immunomodulatory potential. Despite their regenerative potential, ASCs experience replicative senescence during in vitro expansion, resulting in oxidative damage and impaired cellular function. Alpha-lipoic acid (ALA) is a potent antioxidant with reported anti-aging and cytoprotective effects in various cell types. This study aimed to investigate the effects of ALA on proliferation, migration, invasion capacity, and oxidative stress response of primary human ASCs in vitro. Human ASCs were isolated from adipose tissue samples and characterized via flow cytometry and multilineage differentiation. The safe and effective concentration of ALA was determined using WST-1 cell viability assays. Functional evaluations included wound healing and invasion assays. Gene expression of stemness, proliferation, cell cycle, migration, and apoptosis markers was analyzed by qRT-PCR, while MMP2 and MMP9 protein expression was assessed by immunofluorescence. Cellular oxidative stress levels were measured using DCF-DA flow cytometry, and cytotoxicity was evaluated with the LDH release assay. ALA supplementation significantly enhanced ASC proliferation (p < 0.01), migration (p < 0.001), and invasion (p < 0.01). Upregulation of MMP2 (p < 0.001) and MMP9 (p < 0.001) proteins was confirmed by immunofluorescence. ROS levels (p < 0.001) and LDH release (p < 0.001) were markedly reduced in ALA-treated cells under oxidative challenge. The ALA group also maintained higher expression of key proliferation and stemness-associated genes. ALA improves the in vitro expansion and functional properties of ASCs by enhancing proliferative and migratory capacities and mitigating oxidative stress-induced damage. These findings suggest that ALA may serve as a beneficial supplement in ASC-based regenerative applications and long-term culture systems.