title: 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 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.
authors.affiliation: Schepens Eye Research Institute/Massachusetts Eye and Ear, Harvard Medical School, Boston, MA, USA. rakibul.lubikar@gmail.com.|Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway. rakibul.lubikar@gmail.com.|Department of Medical Biochemistry, Oslo University Hospital, Oslo, Norway. rakibul.lubikar@gmail.com.
authors.forename: Rakibul
authors.initials: R
authors.lastname: Islam
authors.affiliation: Department of Ophthalmology, Oslo University Hospital, Oslo, Norway.
authors.forename: Jon Roger
authors.initials: JR
authors.lastname: Eidet
authors.affiliation: Schepens Eye Research Institute/Massachusetts Eye and Ear, Harvard Medical School, Boston, MA, USA.|Faculty of Visual Sciences, University College of Southeast Norway, Kongsberg, Norway.|Department of Ophthalmology, Innlandet Hospital Trust, Elverum, Norway.
authors.forename: Reza A
authors.initials: RA
authors.lastname: Badian
authors.affiliation: Schepens Eye Research Institute/Massachusetts Eye and Ear, Harvard Medical School, Boston, MA, USA.|Department of Medical Biochemistry, Oslo University Hospital, Oslo, Norway.|Faculty of Medicine, University of Oslo, Oslo, Norway.
authors.forename: Marit
authors.initials: M
authors.lastname: Lippestad
authors.affiliation: Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway.
authors.forename: Edward
authors.initials: E
authors.lastname: Messelt
authors.affiliation: Integrative Regenerative Medicine (IGEN) Centre, Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden.
authors.forename: May
authors.identifier: http://orcid.org/0000-0003-1222-6720
authors.initials: M
authors.lastname: Griffith
authors.affiliation: Schepens Eye Research Institute/Massachusetts Eye and Ear, Harvard Medical School, Boston, MA, USA.|Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway.
authors.forename: Darlene A
authors.initials: DA
authors.lastname: Dartt
authors.affiliation: Schepens Eye Research Institute/Massachusetts Eye and Ear, Harvard Medical School, Boston, MA, USA.|Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway.|Department of Medical Biochemistry, Oslo University Hospital, Oslo, Norway.|Department of Ophthalmology, Innlandet Hospital Trust, Elverum, Norway.
authors.forename: Tor Paaske
authors.initials: TP
authors.lastname: Utheim
mesh_terms: D000818:Animals; D015415:Biomarkers; D002448:Cell Adhesion; D049109:Cell Proliferation; D002478:Cells, Cultured; D003470:Culture Media; D004847:Epithelial Cells / Q000166:cytology* / Q000502:physiology*; D007150:Immunohistochemistry; D009061:Mouth Mucosa / Q000166:cytology*; D010641:Phenotype; D051381:Rats
chemical_list: D015415:Biomarkers; D003470:Culture Media