Establishment of Novel Limbus-Derived, Highly Proliferative ABCG2+/ABCB5+ Limbal Epithelial Stem Cell Cultures

Homeostasis and regeneration of corneal epithelia are sustained by limbal epithelial stem cells (LESCs); thus, an LESC deficiency is a major cause of blindness worldwide. Despite the generally promising results of cultivated LESC transplantation, it has been limited by variations in long-term succes...

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Main Authors: Eung Kweon Kim, Ga-Hyun Lee, Boram Lee, Yong-Sun Maeng
Format: Article
Language:English
Published: Wiley 2017-01-01
Series:Stem Cells International
Online Access:http://dx.doi.org/10.1155/2017/7678637
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author Eung Kweon Kim
Ga-Hyun Lee
Boram Lee
Yong-Sun Maeng
author_facet Eung Kweon Kim
Ga-Hyun Lee
Boram Lee
Yong-Sun Maeng
author_sort Eung Kweon Kim
collection DOAJ
description Homeostasis and regeneration of corneal epithelia are sustained by limbal epithelial stem cells (LESCs); thus, an LESC deficiency is a major cause of blindness worldwide. Despite the generally promising results of cultivated LESC transplantation, it has been limited by variations in long-term success rates, the use of xenogeneic and undefined culture components, and a scarcity of donor tissues. In this study, we identified the culture conditions required to expand LESCs in vitro and established human limbus-derived highly proliferative ABCG2+/ABCB5+ double-positive LESCs. These LESCs exhibited the LESC marker profile and differentiated into corneal epithelial cells. In addition, cultured LESCs expressed high levels of the stem cell markers Sox2, Oct4, c-Myc, and Klf4, had high telomerase activity, and had stable, normal genomes. These results suggest that our novel cultivation protocol affects the phenotype and differentiation capacity of LESCs. From the limbus, which contains a heterogenous cell population, we have derived highly proliferative ABCG2+/ABCB5+ double-positive cells with the ability to differentiate into corneal epithelial cells. This study opens a new avenue for investigation of the molecular mechanism of LESC maintenance and expansion in vitro and may impact the treatment of corneal disease, particularly corneal blindness due to an LESC deficiency.
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spelling doaj-art-e5d2b0bd57614fba9ebebc9639afe8282025-02-03T01:02:35ZengWileyStem Cells International1687-966X1687-96782017-01-01201710.1155/2017/76786377678637Establishment of Novel Limbus-Derived, Highly Proliferative ABCG2+/ABCB5+ Limbal Epithelial Stem Cell CulturesEung Kweon Kim0Ga-Hyun Lee1Boram Lee2Yong-Sun Maeng3Department of Ophthalmology, Corneal Dystrophy Research Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaDepartment of Ophthalmology, Corneal Dystrophy Research Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaDepartment of Ophthalmology, Corneal Dystrophy Research Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaDepartment of Ophthalmology, Corneal Dystrophy Research Institute, Yonsei University College of Medicine, Seoul, Republic of KoreaHomeostasis and regeneration of corneal epithelia are sustained by limbal epithelial stem cells (LESCs); thus, an LESC deficiency is a major cause of blindness worldwide. Despite the generally promising results of cultivated LESC transplantation, it has been limited by variations in long-term success rates, the use of xenogeneic and undefined culture components, and a scarcity of donor tissues. In this study, we identified the culture conditions required to expand LESCs in vitro and established human limbus-derived highly proliferative ABCG2+/ABCB5+ double-positive LESCs. These LESCs exhibited the LESC marker profile and differentiated into corneal epithelial cells. In addition, cultured LESCs expressed high levels of the stem cell markers Sox2, Oct4, c-Myc, and Klf4, had high telomerase activity, and had stable, normal genomes. These results suggest that our novel cultivation protocol affects the phenotype and differentiation capacity of LESCs. From the limbus, which contains a heterogenous cell population, we have derived highly proliferative ABCG2+/ABCB5+ double-positive cells with the ability to differentiate into corneal epithelial cells. This study opens a new avenue for investigation of the molecular mechanism of LESC maintenance and expansion in vitro and may impact the treatment of corneal disease, particularly corneal blindness due to an LESC deficiency.http://dx.doi.org/10.1155/2017/7678637
spellingShingle Eung Kweon Kim
Ga-Hyun Lee
Boram Lee
Yong-Sun Maeng
Establishment of Novel Limbus-Derived, Highly Proliferative ABCG2+/ABCB5+ Limbal Epithelial Stem Cell Cultures
Stem Cells International
title Establishment of Novel Limbus-Derived, Highly Proliferative ABCG2+/ABCB5+ Limbal Epithelial Stem Cell Cultures
title_full Establishment of Novel Limbus-Derived, Highly Proliferative ABCG2+/ABCB5+ Limbal Epithelial Stem Cell Cultures
title_fullStr Establishment of Novel Limbus-Derived, Highly Proliferative ABCG2+/ABCB5+ Limbal Epithelial Stem Cell Cultures
title_full_unstemmed Establishment of Novel Limbus-Derived, Highly Proliferative ABCG2+/ABCB5+ Limbal Epithelial Stem Cell Cultures
title_short Establishment of Novel Limbus-Derived, Highly Proliferative ABCG2+/ABCB5+ Limbal Epithelial Stem Cell Cultures
title_sort establishment of novel limbus derived highly proliferative abcg2 abcb5 limbal epithelial stem cell cultures
url http://dx.doi.org/10.1155/2017/7678637
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AT gahyunlee establishmentofnovellimbusderivedhighlyproliferativeabcg2abcb5limbalepithelialstemcellcultures
AT boramlee establishmentofnovellimbusderivedhighlyproliferativeabcg2abcb5limbalepithelialstemcellcultures
AT yongsunmaeng establishmentofnovellimbusderivedhighlyproliferativeabcg2abcb5limbalepithelialstemcellcultures