Hypoxia is a key regulator of limbal epithelial stem cell growth and differentiation

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15 Citationer (Scopus)

Resumé

The aim of this study was to determine whether the growth and differentiation of limbal epithelial stem cell cultures could be controlled through manipulation of the oxygen tension. Limbal epithelial cells were isolated from corneoscleral disks, and cultured using either feeder cells in a growth medium supplemented with serum (3T3 system) or without feeder cells in a dedicated serum-free medium (EpiLife). During the culture, the cells were maintained either at ambient oxygen tension (20%) or at different levels of hypoxia (15, 10, 5, and 2% oxygen). The effect of oxygen on cell growth, progression through cell cycle, colony forming efficiency (CFE), and expression of stem cell (ABCG2 and p63α) and differentiation (CK3) markers was determined throughout the culture period of up to 18 days. Low oxygen levels favored a stem cell phenotype with a lower proliferative rate, high CFE, and a relatively higher expression of ABCG2 and p63α, while higher levels of oxygen led not only to decreased CFE but also to increased proportion of differentiated cells positive for CK3. Hypoxic cultures may thus potentially improve stem cell grafts for cultured limbal epithelial transplantation (CLET).
OriginalsprogEngelsk
TidsskriftStem Cell Research
Vol/bind10
Udgave nummer3
Sider (fra-til)349–360
Antal sider12
ISSN1873-5061
DOI
StatusUdgivet - maj 2013

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Cell Differentiation
Stem Cells
Epithelial Cells
Oxygen
Growth
Feeder Cells
Differentiation Antigens
Serum-Free Culture Media
Hypoxia
Cell Cycle
Cell Culture Techniques
Transplantation
Transplants
Phenotype
Serum

Citer dette

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title = "Hypoxia is a key regulator of limbal epithelial stem cell growth and differentiation",
abstract = "The aim of this study was to determine whether the growth and differentiation of limbal epithelial stem cell cultures could be controlled through manipulation of the oxygen tension. Limbal epithelial cells were isolated from corneoscleral disks, and cultured using either feeder cells in a growth medium supplemented with serum (3T3 system) or without feeder cells in a dedicated serum-free medium (EpiLife). During the culture, the cells were maintained either at ambient oxygen tension (20{\%}) or at different levels of hypoxia (15, 10, 5, and 2{\%} oxygen). The effect of oxygen on cell growth, progression through cell cycle, colony forming efficiency (CFE), and expression of stem cell (ABCG2 and p63α) and differentiation (CK3) markers was determined throughout the culture period of up to 18 days. Low oxygen levels favored a stem cell phenotype with a lower proliferative rate, high CFE, and a relatively higher expression of ABCG2 and p63α, while higher levels of oxygen led not only to decreased CFE but also to increased proportion of differentiated cells positive for CK3. Hypoxic cultures may thus potentially improve stem cell grafts for cultured limbal epithelial transplantation (CLET).",
keywords = "3T3 Cells, Animals, Cell Differentiation, Cell Hypoxia, Cell Proliferation, Cells, Cultured, Coculture Techniques, Feeder Cells, Humans, Keratin-3, Limbus Corneae, Mice, Stem Cells",
author = "S{\o}ndergaard, {Chris Bath} and Sufang Yang and Muttuvelu, {Danson V.} and Trine Fink and Jeppe Emmersen and Henrik Vorum and Hjortdal, {Jesper {\O}stergaard} and Vladimir Zachar",
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Hypoxia is a key regulator of limbal epithelial stem cell growth and differentiation. / Søndergaard, Chris Bath; Yang, Sufang; Muttuvelu, Danson V.; Fink, Trine; Emmersen, Jeppe; Vorum, Henrik; Hjortdal, Jesper Østergaard; Zachar, Vladimir.

I: Stem Cell Research, Bind 10, Nr. 3, 05.2013, s. 349–360.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Hypoxia is a key regulator of limbal epithelial stem cell growth and differentiation

AU - Søndergaard, Chris Bath

AU - Yang, Sufang

AU - Muttuvelu, Danson V.

AU - Fink, Trine

AU - Emmersen, Jeppe

AU - Vorum, Henrik

AU - Hjortdal, Jesper Østergaard

AU - Zachar, Vladimir

PY - 2013/5

Y1 - 2013/5

N2 - The aim of this study was to determine whether the growth and differentiation of limbal epithelial stem cell cultures could be controlled through manipulation of the oxygen tension. Limbal epithelial cells were isolated from corneoscleral disks, and cultured using either feeder cells in a growth medium supplemented with serum (3T3 system) or without feeder cells in a dedicated serum-free medium (EpiLife). During the culture, the cells were maintained either at ambient oxygen tension (20%) or at different levels of hypoxia (15, 10, 5, and 2% oxygen). The effect of oxygen on cell growth, progression through cell cycle, colony forming efficiency (CFE), and expression of stem cell (ABCG2 and p63α) and differentiation (CK3) markers was determined throughout the culture period of up to 18 days. Low oxygen levels favored a stem cell phenotype with a lower proliferative rate, high CFE, and a relatively higher expression of ABCG2 and p63α, while higher levels of oxygen led not only to decreased CFE but also to increased proportion of differentiated cells positive for CK3. Hypoxic cultures may thus potentially improve stem cell grafts for cultured limbal epithelial transplantation (CLET).

AB - The aim of this study was to determine whether the growth and differentiation of limbal epithelial stem cell cultures could be controlled through manipulation of the oxygen tension. Limbal epithelial cells were isolated from corneoscleral disks, and cultured using either feeder cells in a growth medium supplemented with serum (3T3 system) or without feeder cells in a dedicated serum-free medium (EpiLife). During the culture, the cells were maintained either at ambient oxygen tension (20%) or at different levels of hypoxia (15, 10, 5, and 2% oxygen). The effect of oxygen on cell growth, progression through cell cycle, colony forming efficiency (CFE), and expression of stem cell (ABCG2 and p63α) and differentiation (CK3) markers was determined throughout the culture period of up to 18 days. Low oxygen levels favored a stem cell phenotype with a lower proliferative rate, high CFE, and a relatively higher expression of ABCG2 and p63α, while higher levels of oxygen led not only to decreased CFE but also to increased proportion of differentiated cells positive for CK3. Hypoxic cultures may thus potentially improve stem cell grafts for cultured limbal epithelial transplantation (CLET).

KW - 3T3 Cells

KW - Animals

KW - Cell Differentiation

KW - Cell Hypoxia

KW - Cell Proliferation

KW - Cells, Cultured

KW - Coculture Techniques

KW - Feeder Cells

KW - Humans

KW - Keratin-3

KW - Limbus Corneae

KW - Mice

KW - Stem Cells

U2 - 10.1016/j.scr.2013.01.004

DO - 10.1016/j.scr.2013.01.004

M3 - Journal article

VL - 10

SP - 349

EP - 360

JO - Stem Cell Research

JF - Stem Cell Research

SN - 1873-5061

IS - 3

ER -