Effect of retinoic acid on epithelial differentiation and mucin expression in primary human corneal limbal epithelial cells

Sun Woong Kim, KyoungYul Seo, Taiyoun Rhim, Eungkweon Kim

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Purpose: Retinoic acid (RA) is essential for epithelial differentiation and maintenance of the mucous phenotype. This study investigated the effect of RA on corneal epithelial differentiation and mucin expression in a primary human corneal limbal epithelial cell (HCLEC) culture model. Methods: HCLECs were grown in RA-supplemented media at various concentrations (0, 10 -9 to 10 -6 M). Stratified HCLECs were examined using immunohistochemical or immunofluorescent staining for p63, ABCG2, CK3, CK19, and Western blotting for ABCG2 and CK12 to assess differentiation. Ultrastructural morphology was investigated using scanning and transmission electron microscopy. They were incubated with rose bengal dye to examine barrier function. The effects of RA on the expression of MUC1, -4, and -16 were analyzed by immunohistochemistry, quantitative real-time PCR and Western blot analysis. Results: HCLEC grown without RA showed hyperkeratosis, whereas those grown with 10 -8 to 10 -7 M RA induced non-keratinized stratified epithelium with a normal appearance. Under these conditions, p63, ABCG2, CK3, CK19, MUC1, -4, and -16 staining patterns were similar to in vivo limbal epithelium. A higher concentration (10 -6 M) of RA resulted in abnormal differentiation. HCLECs grown with RA were tightly apposed and maintained intact barrier function against dye penetration. In addition, MUC1, -4, and -16 expressions were highly associated with RA concentrations. Conclusions: This study showed that cultured HCLEC could mimic physiologic and functional phenotypes by controlling RA concentrations in medium. Also, our results suggested modulating effect of RA on differentiation and mucin expression in corneal epithelium.

Original languageEnglish
Pages (from-to)33-42
Number of pages10
JournalCurrent Eye Research
Volume37
Issue number1
DOIs
Publication statusPublished - 2012 Jan 1

Fingerprint

Mucins
Tretinoin
Epithelial Cells
Coloring Agents
Epithelium
Western Blotting
Staining and Labeling
Phenotype
Rose Bengal
Corneal Epithelium
Scanning Transmission Electron Microscopy
Real-Time Polymerase Chain Reaction
Cell Culture Techniques
Immunohistochemistry
Maintenance

All Science Journal Classification (ASJC) codes

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

@article{778a08f460c14eeea66802a528a3e01e,
title = "Effect of retinoic acid on epithelial differentiation and mucin expression in primary human corneal limbal epithelial cells",
abstract = "Purpose: Retinoic acid (RA) is essential for epithelial differentiation and maintenance of the mucous phenotype. This study investigated the effect of RA on corneal epithelial differentiation and mucin expression in a primary human corneal limbal epithelial cell (HCLEC) culture model. Methods: HCLECs were grown in RA-supplemented media at various concentrations (0, 10 -9 to 10 -6 M). Stratified HCLECs were examined using immunohistochemical or immunofluorescent staining for p63, ABCG2, CK3, CK19, and Western blotting for ABCG2 and CK12 to assess differentiation. Ultrastructural morphology was investigated using scanning and transmission electron microscopy. They were incubated with rose bengal dye to examine barrier function. The effects of RA on the expression of MUC1, -4, and -16 were analyzed by immunohistochemistry, quantitative real-time PCR and Western blot analysis. Results: HCLEC grown without RA showed hyperkeratosis, whereas those grown with 10 -8 to 10 -7 M RA induced non-keratinized stratified epithelium with a normal appearance. Under these conditions, p63, ABCG2, CK3, CK19, MUC1, -4, and -16 staining patterns were similar to in vivo limbal epithelium. A higher concentration (10 -6 M) of RA resulted in abnormal differentiation. HCLECs grown with RA were tightly apposed and maintained intact barrier function against dye penetration. In addition, MUC1, -4, and -16 expressions were highly associated with RA concentrations. Conclusions: This study showed that cultured HCLEC could mimic physiologic and functional phenotypes by controlling RA concentrations in medium. Also, our results suggested modulating effect of RA on differentiation and mucin expression in corneal epithelium.",
author = "Kim, {Sun Woong} and KyoungYul Seo and Taiyoun Rhim and Eungkweon Kim",
year = "2012",
month = "1",
day = "1",
doi = "10.3109/02713683.2011.620728",
language = "English",
volume = "37",
pages = "33--42",
journal = "Current Eye Research",
issn = "0271-3683",
publisher = "Informa Healthcare",
number = "1",

}

Effect of retinoic acid on epithelial differentiation and mucin expression in primary human corneal limbal epithelial cells. / Kim, Sun Woong; Seo, KyoungYul; Rhim, Taiyoun; Kim, Eungkweon.

In: Current Eye Research, Vol. 37, No. 1, 01.01.2012, p. 33-42.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Effect of retinoic acid on epithelial differentiation and mucin expression in primary human corneal limbal epithelial cells

AU - Kim, Sun Woong

AU - Seo, KyoungYul

AU - Rhim, Taiyoun

AU - Kim, Eungkweon

PY - 2012/1/1

Y1 - 2012/1/1

N2 - Purpose: Retinoic acid (RA) is essential for epithelial differentiation and maintenance of the mucous phenotype. This study investigated the effect of RA on corneal epithelial differentiation and mucin expression in a primary human corneal limbal epithelial cell (HCLEC) culture model. Methods: HCLECs were grown in RA-supplemented media at various concentrations (0, 10 -9 to 10 -6 M). Stratified HCLECs were examined using immunohistochemical or immunofluorescent staining for p63, ABCG2, CK3, CK19, and Western blotting for ABCG2 and CK12 to assess differentiation. Ultrastructural morphology was investigated using scanning and transmission electron microscopy. They were incubated with rose bengal dye to examine barrier function. The effects of RA on the expression of MUC1, -4, and -16 were analyzed by immunohistochemistry, quantitative real-time PCR and Western blot analysis. Results: HCLEC grown without RA showed hyperkeratosis, whereas those grown with 10 -8 to 10 -7 M RA induced non-keratinized stratified epithelium with a normal appearance. Under these conditions, p63, ABCG2, CK3, CK19, MUC1, -4, and -16 staining patterns were similar to in vivo limbal epithelium. A higher concentration (10 -6 M) of RA resulted in abnormal differentiation. HCLECs grown with RA were tightly apposed and maintained intact barrier function against dye penetration. In addition, MUC1, -4, and -16 expressions were highly associated with RA concentrations. Conclusions: This study showed that cultured HCLEC could mimic physiologic and functional phenotypes by controlling RA concentrations in medium. Also, our results suggested modulating effect of RA on differentiation and mucin expression in corneal epithelium.

AB - Purpose: Retinoic acid (RA) is essential for epithelial differentiation and maintenance of the mucous phenotype. This study investigated the effect of RA on corneal epithelial differentiation and mucin expression in a primary human corneal limbal epithelial cell (HCLEC) culture model. Methods: HCLECs were grown in RA-supplemented media at various concentrations (0, 10 -9 to 10 -6 M). Stratified HCLECs were examined using immunohistochemical or immunofluorescent staining for p63, ABCG2, CK3, CK19, and Western blotting for ABCG2 and CK12 to assess differentiation. Ultrastructural morphology was investigated using scanning and transmission electron microscopy. They were incubated with rose bengal dye to examine barrier function. The effects of RA on the expression of MUC1, -4, and -16 were analyzed by immunohistochemistry, quantitative real-time PCR and Western blot analysis. Results: HCLEC grown without RA showed hyperkeratosis, whereas those grown with 10 -8 to 10 -7 M RA induced non-keratinized stratified epithelium with a normal appearance. Under these conditions, p63, ABCG2, CK3, CK19, MUC1, -4, and -16 staining patterns were similar to in vivo limbal epithelium. A higher concentration (10 -6 M) of RA resulted in abnormal differentiation. HCLECs grown with RA were tightly apposed and maintained intact barrier function against dye penetration. In addition, MUC1, -4, and -16 expressions were highly associated with RA concentrations. Conclusions: This study showed that cultured HCLEC could mimic physiologic and functional phenotypes by controlling RA concentrations in medium. Also, our results suggested modulating effect of RA on differentiation and mucin expression in corneal epithelium.

UR - http://www.scopus.com/inward/record.url?scp=83655202850&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=83655202850&partnerID=8YFLogxK

U2 - 10.3109/02713683.2011.620728

DO - 10.3109/02713683.2011.620728

M3 - Article

VL - 37

SP - 33

EP - 42

JO - Current Eye Research

JF - Current Eye Research

SN - 0271-3683

IS - 1

ER -