Tissue Harvesting Site and Culture Medium Affect Attachment, Growth, and Phenotype of Ex Vivo Expanded Oral Mucosal Epithelial Cells

Abstract Transplantation of cultured oral mucosal epithelial cells (OMECs) is a promising treatment strategy for limbal stem cell deficiency. In order to improve the culture method, we investigated the effects of four culture media and tissue harvesting sites on explant attachment, growth, and pheno...

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Autores principales:	Rakibul Islam, Jon Roger Eidet, Reza A. Badian, Marit Lippestad, Edward Messelt, May Griffith, Darlene A. Dartt, Tor Paaske Utheim
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/f767aec0d53645c0b57aefc53d2f67c6
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spelling	oai:doaj.org-article:f767aec0d53645c0b57aefc53d2f67c62021-12-02T12:31:52ZTissue Harvesting Site and Culture Medium Affect Attachment, Growth, and Phenotype of Ex Vivo Expanded Oral Mucosal Epithelial Cells10.1038/s41598-017-00417-z2045-2322https://doaj.org/article/f767aec0d53645c0b57aefc53d2f67c62017-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-00417-zhttps://doaj.org/toc/2045-2322Abstract Transplantation of cultured oral mucosal epithelial cells (OMECs) is a promising treatment strategy for limbal stem cell deficiency. In order to improve the culture method, we investigated the effects of four culture media and tissue harvesting sites on explant attachment, growth, and phenotype of OMECs cultured from Sprague-Dawley rats. Neither choice of media or harvesting site impacted the ability of the explants to attach to the culture well. Dulbecco’s modified Eagle’s medium/Ham’s F12 (DMEM) and Roswell Park Memorial Institute 1640 medium (RPMI) supported the largest cellular outgrowth. Fold outgrowth was superior from LL explants compared to explants from the buccal mucosa (BM), HP, and transition zone of the lower lip (TZ) after six-day culture. Putative stem cell markers were detected in cultures grown in DMEM and RPMI. In DMEM, cells from TZ showed higher colony-forming efficiency than LL, BM, and HP. In contrast to RPMI, DMEM both expressed the putative stem cell marker Bmi-1 and yielded cell colonies. Our data suggest that OMECs from LL and TZ cultured in DMEM give rise to undifferentiated cells with high growth capacity, and hence are the most promising for treatment of limbal stem cell deficiency.Rakibul IslamJon Roger EidetReza A. BadianMarit LippestadEdward MesseltMay GriffithDarlene A. DarttTor Paaske UtheimNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-12 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Rakibul Islam Jon Roger Eidet Reza A. Badian Marit Lippestad Edward Messelt May Griffith Darlene A. Dartt Tor Paaske Utheim Tissue Harvesting Site and Culture Medium Affect Attachment, Growth, and Phenotype of Ex Vivo Expanded Oral Mucosal Epithelial Cells
description	Abstract Transplantation of cultured oral mucosal epithelial cells (OMECs) is a promising treatment strategy for limbal stem cell deficiency. In order to improve the culture method, we investigated the effects of four culture media and tissue harvesting sites on explant attachment, growth, and phenotype of OMECs cultured from Sprague-Dawley rats. Neither choice of media or harvesting site impacted the ability of the explants to attach to the culture well. Dulbecco’s modified Eagle’s medium/Ham’s F12 (DMEM) and Roswell Park Memorial Institute 1640 medium (RPMI) supported the largest cellular outgrowth. Fold outgrowth was superior from LL explants compared to explants from the buccal mucosa (BM), HP, and transition zone of the lower lip (TZ) after six-day culture. Putative stem cell markers were detected in cultures grown in DMEM and RPMI. In DMEM, cells from TZ showed higher colony-forming efficiency than LL, BM, and HP. In contrast to RPMI, DMEM both expressed the putative stem cell marker Bmi-1 and yielded cell colonies. Our data suggest that OMECs from LL and TZ cultured in DMEM give rise to undifferentiated cells with high growth capacity, and hence are the most promising for treatment of limbal stem cell deficiency.
format	article
author	Rakibul Islam Jon Roger Eidet Reza A. Badian Marit Lippestad Edward Messelt May Griffith Darlene A. Dartt Tor Paaske Utheim
author_facet	Rakibul Islam Jon Roger Eidet Reza A. Badian Marit Lippestad Edward Messelt May Griffith Darlene A. Dartt Tor Paaske Utheim
author_sort	Rakibul Islam
title	Tissue Harvesting Site and Culture Medium Affect Attachment, Growth, and Phenotype of Ex Vivo Expanded Oral Mucosal Epithelial Cells
title_short	Tissue Harvesting Site and Culture Medium Affect Attachment, Growth, and Phenotype of Ex Vivo Expanded Oral Mucosal Epithelial Cells
title_full	Tissue Harvesting Site and Culture Medium Affect Attachment, Growth, and Phenotype of Ex Vivo Expanded Oral Mucosal Epithelial Cells
title_fullStr	Tissue Harvesting Site and Culture Medium Affect Attachment, Growth, and Phenotype of Ex Vivo Expanded Oral Mucosal Epithelial Cells
title_full_unstemmed	Tissue Harvesting Site and Culture Medium Affect Attachment, Growth, and Phenotype of Ex Vivo Expanded Oral Mucosal Epithelial Cells
title_sort	tissue harvesting site and culture medium affect attachment, growth, and phenotype of ex vivo expanded oral mucosal epithelial cells
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/f767aec0d53645c0b57aefc53d2f67c6
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Tissue Harvesting Site and Culture Medium Affect Attachment, Growth, and Phenotype of Ex Vivo Expanded Oral Mucosal Epithelial Cells

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