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Human corneal stromal stem cells support limbal epithelial cells cultured on RAFT tissue equivalents
Human limbal epithelial cells (HLE) and corneal stromal stem cells (CSSC) reside in close proximity in vivo in the corneal limbal stem cell niche. However, HLE are typically cultured in vitro without supporting niche cells. Here, we re-create the cell-cell juxtaposition of the native environment in...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4632025/ https://www.ncbi.nlm.nih.gov/pubmed/26531048 http://dx.doi.org/10.1038/srep16186 |
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author | Kureshi, Alvena K Dziasko, Marc Funderburgh, James L Daniels, Julie T |
author_facet | Kureshi, Alvena K Dziasko, Marc Funderburgh, James L Daniels, Julie T |
author_sort | Kureshi, Alvena K |
collection | PubMed |
description | Human limbal epithelial cells (HLE) and corneal stromal stem cells (CSSC) reside in close proximity in vivo in the corneal limbal stem cell niche. However, HLE are typically cultured in vitro without supporting niche cells. Here, we re-create the cell-cell juxtaposition of the native environment in vitro, to provide a tool for investigation of epithelial-stromal cell interactions and to optimize HLE culture conditions for potential therapeutic application. RAFT (Real Architecture For 3D Tissue) tissue equivalents (TEs) were used as a 3-dimensional substrate for co-culturing HLE and CSSC. Our results demonstrate that a monolayer of HLE that maintained expression of p63α, ABCB5, CK8 and CK15 (HLE markers), formed on the surface of RAFT TEs within 13 days of culture. CSSC remained in close proximity to HLE and maintained expression of mesenchymal stem cell markers. This simple technique has a short preparation time of only 15 days with the onset of HLE layering and differentiation observed. Furthermore, co-cultivation of HLE with another niche cell type (CSSC) directly on RAFT TEs, eliminates the requirement for animal-derived feeder cells. RAFT TEs may be useful for future therapeutic delivery of multiple cell types to restore the limbal niche following ocular surface injury or disease. |
format | Online Article Text |
id | pubmed-4632025 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-46320252015-12-07 Human corneal stromal stem cells support limbal epithelial cells cultured on RAFT tissue equivalents Kureshi, Alvena K Dziasko, Marc Funderburgh, James L Daniels, Julie T Sci Rep Article Human limbal epithelial cells (HLE) and corneal stromal stem cells (CSSC) reside in close proximity in vivo in the corneal limbal stem cell niche. However, HLE are typically cultured in vitro without supporting niche cells. Here, we re-create the cell-cell juxtaposition of the native environment in vitro, to provide a tool for investigation of epithelial-stromal cell interactions and to optimize HLE culture conditions for potential therapeutic application. RAFT (Real Architecture For 3D Tissue) tissue equivalents (TEs) were used as a 3-dimensional substrate for co-culturing HLE and CSSC. Our results demonstrate that a monolayer of HLE that maintained expression of p63α, ABCB5, CK8 and CK15 (HLE markers), formed on the surface of RAFT TEs within 13 days of culture. CSSC remained in close proximity to HLE and maintained expression of mesenchymal stem cell markers. This simple technique has a short preparation time of only 15 days with the onset of HLE layering and differentiation observed. Furthermore, co-cultivation of HLE with another niche cell type (CSSC) directly on RAFT TEs, eliminates the requirement for animal-derived feeder cells. RAFT TEs may be useful for future therapeutic delivery of multiple cell types to restore the limbal niche following ocular surface injury or disease. Nature Publishing Group 2015-11-04 /pmc/articles/PMC4632025/ /pubmed/26531048 http://dx.doi.org/10.1038/srep16186 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Kureshi, Alvena K Dziasko, Marc Funderburgh, James L Daniels, Julie T Human corneal stromal stem cells support limbal epithelial cells cultured on RAFT tissue equivalents |
title | Human corneal stromal stem cells support limbal epithelial cells cultured on RAFT tissue equivalents |
title_full | Human corneal stromal stem cells support limbal epithelial cells cultured on RAFT tissue equivalents |
title_fullStr | Human corneal stromal stem cells support limbal epithelial cells cultured on RAFT tissue equivalents |
title_full_unstemmed | Human corneal stromal stem cells support limbal epithelial cells cultured on RAFT tissue equivalents |
title_short | Human corneal stromal stem cells support limbal epithelial cells cultured on RAFT tissue equivalents |
title_sort | human corneal stromal stem cells support limbal epithelial cells cultured on raft tissue equivalents |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4632025/ https://www.ncbi.nlm.nih.gov/pubmed/26531048 http://dx.doi.org/10.1038/srep16186 |
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