Limbal epithelial stem cell sheets from young donors have better regenerative potential

Area of Science:

• Ophthalmology
• Stem Cell Biology
• Regenerative Medicine

Background:

• Limbal epithelial stem cells (LESCs) are crucial for corneal regeneration.
• Donor age is a potential factor influencing the quality and regenerative capacity of LESC sheets.
• Understanding age-related changes in LESCs is vital for optimizing cell-based therapies.

Purpose of the Study:

• To investigate the impact of donor age on the stemness and regenerative potential of human limbal epithelial stem cell sheets.
• To evaluate key stemness markers, proliferation, and differentiation capacity in relation to donor age groups.

Main Methods:

• Human limbal explants from different age groups (≤45, 45-65, >65 years) were cultured on amniotic membrane scaffolds.
• Evaluation of limbal epithelial sheet size, stem/progenitor cell markers (p63α, ABCG2), proliferation (Ki67), and differentiation (CK12) markers.
• Assessment of colony formation efficiency (CFE) and storage stability using flow cytometry (JC-1, p63α, PANCK).

Main Results:

• Limbal epithelial stem cell sheets from younger donors (≤45 years) exhibited significantly larger outgrowth areas and higher expression of stemness markers (p63α, ABCG2).
• Donors ≤45 years showed increased clonogenic capacity (CFE) and better resistance to cold storage compared to older donors (>65 years).
• Aged donors (>65 years) displayed reduced stemness marker expression and lower regenerative potential.

Conclusions:

• Donor age is a critical determinant of limbal epithelial stem cell sheet quality and regenerative capacity.
• Donors younger than 65 years are preferable for generating limbal epithelial stem cell sheets with high regeneration potential.
• These findings have implications for donor selection in ocular surface reconstruction and regenerative medicine strategies.