The effect of N-acetyl-cysteine on limbal stem cell senescence cultivated on the human amniotic membrane

Document Type : Original Article

Authors

1 Cellular and Molecular Research Center, Sabzevar University of Medical Sciences, Sabzevar, Iran.

2 Department of Clinical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

10.30468/beyhagh.2026.1855

Abstract

Introduction:
In limbal stem cell deficiency disease, limbal stem cells (LSCs) are unable to replace lost epithelial cells. This causes reduced eyesight and blindness. In recent years, the use of cultured LSCs for transplantation has been considered, but their proliferative potency declines during culture due to a premature-stress-induced senescence. For this purpose, the antioxidant effect of N-acetyl-cysteine on the aging process of cultured LSCs was surveyed.
Methods:
LSCs were cultivated on the amniotic membrane in three groups: the test group treated with N-acetyl-cysteine, the positive control group treated with H2O2, and the untreated group as a control. The presence and proliferation of LSCs during culture were assessed by hematoxylin-eosin staining and the expression of ΔNP63α specific marker by immunofluorescent assay. The effect of N-acetyl-cysteine on the senescence process of expanded LSCs was evaluated by measuring beta-galactosidase activity.
Results:
The presence of cells and the proliferation of LSCs during culture were confirmed by hematoxylin-eosin and immunofluorescent staining, respectively. Cells treated with N-acetyl-cysteine in short-term treatment and long-term treatment significantly presented a lower level of beta-galactosidase, which confirms the effective role of N-acetyl-cysteine in delaying the senescence of cultured LSCs.
Conclusion:
Treating LSCs with N-acetyl-cysteine during the culture process could be effective in delaying the senescence of LSCs, which could be valuable for long-term cornea restoration.

Keywords

Beyhagh
Volume 31, Issue 1
June 2026
Pages 61-72

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