Cox, Eric

De Rooster, Hilde

Pelst, Michael

Wallaeys, Charlotte

English

Ghent University Academic Bibliography

96h in cultureα6+ cellsα6- cellsHypothesis of the studyThe surface marker α6 (CD49f) can be used to select canineprimary oral epithelial cells derived from biopsies in order toestablish a cell culture of purified oral keratinocytes.BackgroundApart from forming an important physical barrier, the oral epithelium isan immunologically active first contact zone to a variety of non-selfantigens. For skin keratinocytes such an immunologic response hasbeen shown to influence subepithelial cells, such as antigen-presentingcells, with a potential impact on the development of antigen-specificdiseases like allergies.1 To objectively study these immune responses inoral keratinocytes, a homogenous cell culture should be generated.Human oral keratinocyte stem cells can be selected using the integrinsubunit α6,2 a surface marker which is also expressed by canineepidermal keratinocytes.3 In this study we evaluated whether α6 can beused to characterize and positively select basal keratinocytes in a cellpopulation derived from a biopsy of the canine oral mucosa.MethodsOral biopsies were collected from dogs that were euthanized at thefaculty clinic for causes independent of this study. One half of eachbiopsy was snap-frozen in liquid nitrogen and cryosections were stainedfor α6. The other half was used to generate a purified population ofepithelial cells using the following procedure:References1Asahina, R. and Maeda, S. (2017). A review of the roles of keratinocyte‐derived cytokines and chemokines in the pathogenesis of atopic dermatitis in humans and dogs. In: Torres, S. M. and Roudebush, P. (editors) Advances in Veterinary Dermatology, vol. 8, Wiley-Blackwell, Hoboken, 17-25. 2Calenic, B., Ishkitiev, N., Yaegaki, K., Imai, T., Kumazawa, Y., Nasu, M., & Hirata, T. (2010). Magnetic separation and characterization of keratinocyte stem cells from human gingiva. Journal of periodontal research, 45(6), 703-708.3Shibata, S., Maeda, S., Tsuchida, H., & Fukata, T. (2008). Phenotypic analysis for a cell line of canine epidermal keratinocytes. Journal of Veterinary Medical Science, 70(8), 853-855.ResultsCryosection of the canine sublingual mucosa stained for α6Sorting of α6- and α6+ canine oral mucosal cellsConclusion: The integrin subunit α6 can be used to positively select canineoral keratinocytes. The establishment of a purified cell culturecan facilitate our understanding of the immunologic signallingmechanisms generated by oral keratinocytes. It will allow tomap the cell mediators that are produced by these cells afterexposure to a variety of immunologically active molecules suchas toll-like receptor ligands and allergens. Knowledge of therepertoire of mediators produced by these cells will providefurther opportunities to study how to modulate this immuneresponse for in vivo applications such as desensitizationstrategies for allergies or the oral administration of vaccines.PELST M.a, WALLAEYS C.a, DE ROOSTER H.b, COX E.aaLaboratory of  veterinary immunology, faculty of veterinary medicine, Ghent university, Merelbeke, Belgium bSmall animal department, faculty of veterinary medicine, Ghent university, Merelbeke, Belgium ESTABLISHMENT OF A CELL CULTURE OF CANINE ORAL KERATINOCYTES: ALPHA6 AS A MARKER FOR POSITIVE SELECTIONContactEmail: Michael.Pelst@ugent.beWebsite: http://www.vetimmunology.ugent.be/Universiteit Gent@ugentGhent UniversityDispase+ TrypsinAnti-α6 antibodies+ + ViabilitymarkerSortViable α6- cellsViable α6+ cellsUpper layers of the oralepitheliumBasal layer of the oral epithelium expressing α6 for attachment to basement membraneSubepithelial matrix withstromal cellsEpitheliumSubepithelialtissueBiopsy

Establishment of a cell culture of canine oral keratinocytes : alpha6 as a marker for positive selection

https://biblio.ugent.be/publication/8631206/file/8631208.pdf

Establishment of a cell culture of canine oral keratinocytes : alpha6 as a marker for positive selection

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Ghent University Academic Bibliography