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

• Published in: Scientific reports 2022-08, Vol.12 (1), p.14191-14191, Article 14191
• Main Authors: Yang, Soonwon, Lee, Hyun Jung, Shin, Soojung, Park, In Yang, Chung, So-Hyang
• Format: Article
• Language: English
• Subjects: 631/532; 692/699/3161; Age; Age groups; Amniotic membrane; Cadavers; Cornea; Explants; Flow cytometry; Humanities and Social Sciences; multidisciplinary; Progenitor cells; Science; Science (multidisciplinary); Stem cells
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-022-17821-9

To investigate the stemness of limbal epithelial stem cell sheets in relation to the donor’s age. Human limbal explants from cadaveric donors were set on human amniotic membrane scaffolds with the xeno-free medium. We evaluated limbal epithelial sheet size, expression of stem/progenitor cell markers, and colony formation efficiency from donors of different age groups (age ≤ 45, age 45–65, and age > 65). Expression of the proliferation marker Ki67, stem/progenitor cell markers p63α and ABCG2, cornea specific marker PANCK, and differentiation marker CK12 were evaluated. To determine the effect of donor age on the storage period of limbal explant sheets, the limbal explant outgrowth sheets were stored in 4 °C for 2 days and analyzed for JC-1, p63α, and PANCK with FACS on each day. From days 6 to 12, the outgrowth area of the limbal epithelial stem cell sheet was significantly larger in the age ≤ 45 groups (296 ± 54.7 mm 2 , day 9) compared to the other two age groups [age 45–65 group (278 ± 62.6 mm 2 ), age > 65 group (257 ± 44.0 mm 2 ), day 9] (p  65) showed lower expression of stem/progenitor cell markers p63α and ABCG2 and decreased CFE compared to the other two groups. There were significantly more p63α+ cells in outgrowth cells in the age ≤ 45 group (18.2 ± 3.6%) compared to the age > 65 group (14.1 ± 4.6%; p  65 groups (25.7 ± 7.1%, p