Abstract
Primary human oral epithelial cells are readily available and have been recently employed for tissue engineering. These cells are currently being widely utilized in multiple research efforts, ranging from the study of oral biology, mucosal immunity, and carcinogenesis to stem cell biology and tissue engineering. This chapter describes step-by-step protocols for the successful isolation and culture of human oral epithelial cells and fibroblasts, and techniques for their use in two-dimensional and three-dimensional culture systems. The described methods will enable to generate reconstituted tissues that resemble epithelial like structures in vitro, which can recapitulate some of the key features of the oral epithelium in vivo.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Nanci A, Ten Cate R (2008) Ten Cate’s oral histology: development, structure, and function. Mosby Elsevier Health Sciences, St. Louis, MO
Wertz PW, Squier CA (1991) Cellular and molecular basis of barrier function in oral epithelium. Crit Rev Ther Drug Carrier Syst 8:237–269
Castilho RM, Squarize CH, Leelahavanichkul K, Zheng Y, Bugge T, Gutkind JS (2010) Rac1 is required for epithelial stem cell function during dermal and oral mucosal wound healing but not for tissue homeostasis in mice. PLoS One 5:e10503
Xu X, Lyle S, Liu Y, Solky B, Cotsarelis G (2003) Differential expression of cyclin D1 in the human hair follicle. Am J Pathol 163:969–978
Levy V, Lindon C, Zheng Y, Harfe BD, Morgan BA (2007) Epidermal stem cells arise from the hair follicle after wounding. FASEB J 21:1358–1366
Langton AK, Herrick SE, Headon DJ (2008) An extended epidermal response heals cutaneous wounds in the absence of a hair follicle stem cell contribution. J Invest Dermatol 128:1311–1318
Ito M, Liu Y, Yang Z, Nguyen J, Liang F, Morris RJ, Cotsarelis G (2005) Stem cells in the hair follicle bulge contribute to wound repair but not to homeostasis of the epidermis. Nat Med 11:1351–1354
Levy V, Lindon C, Harfe BD, Morgan BA (2005) Distinct stem cell populations regenerate the follicle and interfollicular epidermis. Dev Cell 9:855–861
Grossman N, Slovik Y, Bodner L (2004) Effect of donor age on cultivation of human oral mucosal keratinocytes. Arch Gerontol Geriatr 38:114–122
Kedjarune U, Pongprerachok S, Arpornmaeklong P, Ungkusonmongkhon K (2001) Culturing primary human gingival epithelial cells: comparison of two isolation techniques. J Craniomaxillofac Surg 29:224–231
Sundqvist K, Liu Y, Arvidson K, Ormstad K, Nilsson L, Toftgard R, Grafstrom RC (1991) Growth regulation of serum-free cultures of epithelial cells from normal human buccal mucosa. In Vitro Cell Dev Biol 27A:562–568
Neugebauer P, Bonnekoh B, Wevers A, Michel O, Mahrle G, Krieg T, Stennert E (1996) Human keratinocyte culture from the peritonsillar mucosa. Eur Arch Otorhinolaryngol 253:245–251
Rickert D, Franke RP, Fernandez CA, Kilroy S, Yan L, Moses MA (2007) Establishment and biochemical characterization of primary cells of the upper aerodigestive tract. Clin Hemorheol Microcirc 36:47–64
Meier F, Nesbit M, Hsu MY, Martin B, Van BP, Elder DE, Schaumburg-Lever G, Garbe C, Walz TM, Donatien P et al (2000) Human melanoma progression in skin reconstructs: biological significance of bFGF. Am J Pathol 156:193–200
Margulis A, Zhang W, Garlick JA (2005) In vitro fabrication of engineered human skin. Methods Mol Biol 289:61–70
Formanek M, Millesi W, Willheim M, Scheiner O, Kornfehl J (1996) Optimized growth medium for primary culture of human oral keratinocytes. Int J Oral Maxillofac Surg 25:157–160
Schaller M, Weindl G (2009) Models of oral and vaginal candidiasis based on in vitro reconstituted human epithelia for the study of host-pathogen interactions. Methods Mol Biol 470:327–345
Tomakidi P, Schuster G, Breitkreutz D, Kohl A, Ottl P, Komposch G (2000) Organotypic cultures of gingival cells: an epithelial model to assess putative local effects of orthodontic plate and occlusal splint materials under more tissue-like conditions. Biomaterials 21:1549–1559
Egles C, Garlick JA, Shamis Y (2010) Three-dimensional human tissue models of wounded skin. Methods Mol Biol 585:345–359
Nystrom ML, Thomas GJ, Stone M, MacKenzie IC, Hart IR, Marshall JF (2005) Development of a quantitative method to analyse tumour cell invasion in organotypic culture. J Pathol 205:468–475
Yoo GH, Washington J, Piechocki M, Ensley J, Shibuya T, Oda D, Wei WZ (2000) Progression of head and neck cancer in an in vitro model. Arch Otolaryngol Head Neck Surg 126:1313–1318
Funding
This research was supported by the Intramural Research Program of the National Institutes of Health, National Institute of Dental and Craniofacial Research, project Z01DE00558. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Leelahavanichkul, K., Gutkind, J.S. (2012). Oral and Pharyngeal Epithelial Keratinocyte Culture. In: Randell, S., Fulcher, M. (eds) Epithelial Cell Culture Protocols. Methods in Molecular Biology, vol 945. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-125-7_5
Download citation
DOI: https://doi.org/10.1007/978-1-62703-125-7_5
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-124-0
Online ISBN: 978-1-62703-125-7
eBook Packages: Springer Protocols