Abstract
Limbal epithelial stem cells (LESCs) maintain the corneal epithelium throughout life and are crucial for both corneal integrity and vision. In this study, LESCs were expanded in either a culture system using 3T3 feeder cells in growth medium supplemented with serum, or in a culture system without feeder cells using commercially available serum-free medium (EpiLife). Cells were maintained at an ambient oxygen concentration of 20% or at various levels of hypoxia (15%, 10%, 5%, and 2%) throughout the period of expansion. The effect of ambient oxygen concentration on growth, cell cycle, colony forming efficiency (CFE), and expression of stem cell markers ABCG2 and p63α and differentiation marker CK3 were determined at different time points. Low oxygen levels were found to maintain a stem cell phenotype with low proliferative rate, high CFE, and high expression of ABCG2 and p63α as well as low expression of CK3. The relation between degree of differentiation and ambient oxygen concentration in the culture system seems to mirror the natural environment of the limbal niche. Hypoxic culture could therefore potentially improve stem cell grafts for cultured limbal epithelial transplantation (CLET).
| Original language | English |
|---|---|
| Title of host publication | The Stem Cell Microenvironment and Its Role in Regenerative Medicine and Cancer Pathogenesis |
| Number of pages | 12 |
| Publisher | River Publishers |
| Publication date | 1 Feb 2017 |
| Pages | 1-12 |
| ISBN (Print) | 9788793379930 |
| ISBN (Electronic) | 9788793519008 |
| Publication status | Published - 1 Feb 2017 |
Keywords
- Adult stem cells
- Cell hypoxia
- Limbus cornea
- Primary cell culture
- Regenerative medicine