Selective expansion of limbal epithelial stem cells in culture

Chris Bath*, Sufang Yang, Danson Muttuvelu, Trine Fink, Jeppe Emmersen, Henrik Vorum, Jesper Hjortdal, Vladimir Zachar

*Corresponding author af dette arbejde

Publikation: Bidrag til bog/antologi/rapport/proceedingBidrag til bog/antologiForskningpeer review

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).

OriginalsprogEngelsk
TitelThe Stem Cell Microenvironment and Its Role in Regenerative Medicine and Cancer Pathogenesis
Antal sider12
ForlagRiver Publishers
Publikationsdato1 feb. 2017
Sider1-12
ISBN (Trykt)9788793379930
ISBN (Elektronisk)9788793519008
StatusUdgivet - 1 feb. 2017

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