Phenotypic change and induction of cytokeratin expression during in vitro culture of corneal stromal cells
Purpose: Cells of the corneal epithelium and stroma can be distinguished in vivo by different intermediate filaments, cytokeratins for corneal epithelial cells (CEC) and vimentin for keratocytes. Isolated and cultured keratocytes change phenotype, losing expression of keratocyte markers and gaining...
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| Format: | Article |
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ARVO
2015
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| Online Access: | https://eprints.nottingham.ac.uk/31252/ |
| _version_ | 1848794160153231360 |
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| author | Sidney, Laura E. McIntosh, Owen D. Hopkinson, Andrew |
| author_facet | Sidney, Laura E. McIntosh, Owen D. Hopkinson, Andrew |
| author_sort | Sidney, Laura E. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Purpose: Cells of the corneal epithelium and stroma can be distinguished in vivo by different intermediate filaments, cytokeratins for corneal epithelial cells (CEC) and vimentin for keratocytes. Isolated and cultured keratocytes change phenotype, losing expression of keratocyte markers and gaining markers associated with mesenchymal stromal cells (MSC). This study investigates this change in phenotype in relation to intermediate filament expression in cultured corneal stromal cells (CSC) compared to CEC.
Methods: Expression of epithelial markers (CK3, CK12, CK19, pan cytokeratin, E-cadherin), keratocyte markers (CD34, vimentin) and MSC markers (CD73, CD90 and CD105) were compared in CEC and CSC by immunocytochemistry and RT-qPCR. Expression was evaluated at different stages of CSC culture and compared to another stromal cell type, extracted from Wharton’s jelly (WJ-MSC).
Results: In vivo keratocytes did not express cytokeratins. However, cultured CSC expressed epithelial-associated CK3, CK12 and CK19 but other cytokeratins. Expression of cytokeratins increased as CSC were passaged and decreased as CSC were induced to become quiescent. Comparatively, WJ-MSC, expressed lower levels of CK3, CK12 and CK19, but also stained for pan cytokeratin and expressed KRT5.
Conclusions: Cultured CSC undergo phenotypic change during culture, expressing specific cytokeratin filaments normally associated with CEC. Cytokeratin expression begins as cells are cultured on plastic and increases with passage. This discovery may influence the way that differences are discerned between cultured CEC and CSC. Investigators need to be aware that the expression of cytokeratins does not necessarily represent epithelial contamination, and that CEC and CSC may be more related than previously recognised. |
| first_indexed | 2025-11-14T19:11:46Z |
| format | Article |
| id | nottingham-31252 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:11:46Z |
| publishDate | 2015 |
| publisher | ARVO |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-312522020-05-04T20:06:39Z https://eprints.nottingham.ac.uk/31252/ Phenotypic change and induction of cytokeratin expression during in vitro culture of corneal stromal cells Sidney, Laura E. McIntosh, Owen D. Hopkinson, Andrew Purpose: Cells of the corneal epithelium and stroma can be distinguished in vivo by different intermediate filaments, cytokeratins for corneal epithelial cells (CEC) and vimentin for keratocytes. Isolated and cultured keratocytes change phenotype, losing expression of keratocyte markers and gaining markers associated with mesenchymal stromal cells (MSC). This study investigates this change in phenotype in relation to intermediate filament expression in cultured corneal stromal cells (CSC) compared to CEC. Methods: Expression of epithelial markers (CK3, CK12, CK19, pan cytokeratin, E-cadherin), keratocyte markers (CD34, vimentin) and MSC markers (CD73, CD90 and CD105) were compared in CEC and CSC by immunocytochemistry and RT-qPCR. Expression was evaluated at different stages of CSC culture and compared to another stromal cell type, extracted from Wharton’s jelly (WJ-MSC). Results: In vivo keratocytes did not express cytokeratins. However, cultured CSC expressed epithelial-associated CK3, CK12 and CK19 but other cytokeratins. Expression of cytokeratins increased as CSC were passaged and decreased as CSC were induced to become quiescent. Comparatively, WJ-MSC, expressed lower levels of CK3, CK12 and CK19, but also stained for pan cytokeratin and expressed KRT5. Conclusions: Cultured CSC undergo phenotypic change during culture, expressing specific cytokeratin filaments normally associated with CEC. Cytokeratin expression begins as cells are cultured on plastic and increases with passage. This discovery may influence the way that differences are discerned between cultured CEC and CSC. Investigators need to be aware that the expression of cytokeratins does not necessarily represent epithelial contamination, and that CEC and CSC may be more related than previously recognised. ARVO 2015-11 Article PeerReviewed Sidney, Laura E., McIntosh, Owen D. and Hopkinson, Andrew (2015) Phenotypic change and induction of cytokeratin expression during in vitro culture of corneal stromal cells. Investigative Ophthalmology and Visual Science, 56 (12). pp. 7225-7235. ISSN 1552-5783 Cornea Corneal stromal cells Cytokeratins Corneal epithelial cells Mesenchymal stem cells http://iovs.arvojournals.org/article.aspx?articleid=2469548 doi:10.1167/iovs.15-17810 doi:10.1167/iovs.15-17810 |
| spellingShingle | Cornea Corneal stromal cells Cytokeratins Corneal epithelial cells Mesenchymal stem cells Sidney, Laura E. McIntosh, Owen D. Hopkinson, Andrew Phenotypic change and induction of cytokeratin expression during in vitro culture of corneal stromal cells |
| title | Phenotypic change and induction of cytokeratin expression during in vitro culture of corneal stromal cells |
| title_full | Phenotypic change and induction of cytokeratin expression during in vitro culture of corneal stromal cells |
| title_fullStr | Phenotypic change and induction of cytokeratin expression during in vitro culture of corneal stromal cells |
| title_full_unstemmed | Phenotypic change and induction of cytokeratin expression during in vitro culture of corneal stromal cells |
| title_short | Phenotypic change and induction of cytokeratin expression during in vitro culture of corneal stromal cells |
| title_sort | phenotypic change and induction of cytokeratin expression during in vitro culture of corneal stromal cells |
| topic | Cornea Corneal stromal cells Cytokeratins Corneal epithelial cells Mesenchymal stem cells |
| url | https://eprints.nottingham.ac.uk/31252/ https://eprints.nottingham.ac.uk/31252/ https://eprints.nottingham.ac.uk/31252/ |