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Gursoy UK, Gursoy M, Könönen E, et al. Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model. Cytotechnology. 2016;68(6):2345-2354doi: 10.1007/s10616-016-0029-4.
Gursoy, U. K., Gursoy, M., Könönen, E., Sintim, H. O., Uitto, V. J., & Syrjänen, S. (2016). Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model. Cytotechnology, 68(6), 2345-2354. https://doi.org/10.1007/s10616-016-0029-4
Gursoy, Ulvi K, et al. "Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model." Cytotechnology vol. 68,6 (2016): 2345-2354. doi: https://doi.org/10.1007/s10616-016-0029-4
Gursoy UK, Gursoy M, Könönen E, Sintim HO, Uitto VJ, Syrjänen S. Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model. Cytotechnology. 2016 Dec;68(6):2345-2354. doi: 10.1007/s10616-016-0029-4. Epub 2016 Oct 17. PMID: 27752840; PMCID: PMC5101304.
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Cytotechnology. 2016 Dec;68(6):2345-2354. doi: 10.1007/s10616-016-0029-4. Epub 2016 Oct 17.

Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model.

Cytotechnology

Ulvi K Gursoy, Mervi Gursoy, Eija Könönen, Herman O Sintim, Veli-Jukka Uitto, Stina Syrjänen

Affiliations

  1. Department of Periodontology, Institute of Dentistry, University of Turku, Lemminkäisenkatu 2, 20520, Turku, Finland. [email protected].
  2. Department of Periodontology, Institute of Dentistry, University of Turku, Lemminkäisenkatu 2, 20520, Turku, Finland.
  3. Oral Health Care, Welfare Division, Turku, Finland.
  4. Department of Chemistry and Purdue Institute for Drug Discovery and Purdue Institute of Inflammation, Immunology and Infectious Disease, Purdue University, West Lafayette, IN, USA.
  5. Department of Oral and Maxillofacial Diseases, University of Helsinki, Helsinki, Finland.
  6. Department of Oral Pathology, Institute of Dentistry, University of Turku, Turku, Finland.

PMID: 27752840 PMCID: PMC5101304 DOI: 10.1007/s10616-016-0029-4

Abstract

In construction of epithelial cells as multilayers, the cells are grown submerged to confluence on fibroblast-embedded collagen gels and, then, lifted to air to promote their stratification. We recently demonstrated that gingival epithelial cells form uniform monolayers on semi-permeable nitrocellulose membranes, supported with a semi-solid growth medium, which allows the cells to grow at an air-liquid-solid interface from the beginning of the culturing protocol. In this study, the aim was to further develop our previous model to form a multilayered gingival epithelial culture model. Two different epithelial cell lines (HaCaT from skin and HMK from gingiva) were used in all experiments. Both cell lines were grown first as monolayers for 3 days. After that, keratinocytes were trypsinized, counted and seeded on a sterile semi-permeable nitrocellulose membrane placed on the top of a semi-solid growth medium, forming an air-liquid-solid interface for the cells to grow. At days 1, 4, and 7, epithelial cells were fixed, embedded in paraffin, and sectioned for routine Hematoxylin-Eosin staining and immunohistochemistry for cytokeratin (Ck). At day 1, HMK cells grew as monolayers, while HaCaT cells stratified forming an epithelium with two to three layers. At day 4, a stratified epithelium in the HMK model had four to five layers and its proliferation continued up to day 7. HaCaT cells formed a dense and weakly proliferating epithelium with three to four layers of stratification at day 4 but the proliferation disappeared at day 7. At all days, both models were strongly positive for Ck5, Ck7, and Ck 19, and weakly positive for Ck10. Gingival epithelial cells stratify successfully on semi-permeable nitrocellulose membranes, supported with a semi-solid growth medium. This technique allows researchers to construct uniform gingival epithelial cell multilayers at an air-liquid-solid interface, without using collagen gels, resulting in a more reproducible method.

Keywords: Epithelium; Keratinocytes; Organotypic

References

  1. Nat Protoc. 2006;1(4):2012-8 - PubMed
  2. Periodontol 2000. 1997 Feb;13:91-120 - PubMed
  3. J Dent Res. 1986 Nov;65(11):1326-31 - PubMed
  4. J Periodontol. 2012 Feb;83(2):242-7 - PubMed
  5. J Cell Physiol. 2015 Jan;230(1):16-26 - PubMed
  6. BMC Oral Health. 2014 Apr 03;14:30 - PubMed
  7. Acta Oncol. 1997;36(1):13-6 - PubMed
  8. J Dent Res. 2012 Jul;91(7):642-50 - PubMed
  9. J Periodontol. 2016 Feb;87(2):193-202 - PubMed
  10. Differentiation. 1984;26(2):154-69 - PubMed
  11. Differentiation. 1989 Mar;40(1):42-54 - PubMed
  12. PLoS One. 2012;7(6):e38599 - PubMed
  13. J Periodontal Res. 1991 Nov;26(6):468-78 - PubMed
  14. Exp Cell Res. 1997 Aug 25;235(1):180-7 - PubMed
  15. Arch Oral Biol. 2014 May;59(5):461-9 - PubMed
  16. Microb Pathog. 2011 Jun;50(6):278-85 - PubMed
  17. J Dairy Sci. 1978 Jun;61(6):729-32 - PubMed
  18. J Periodontol. 2010 Jul;81(7):1084-91 - PubMed
  19. J Oral Microbiol. 2012;4:null - PubMed
  20. J Periodontol. 2002 Jan;73(1):86-93 - PubMed
  21. J Dent Res. 2007 Feb;86(2):115-24 - PubMed
  22. Cell. 1975 Nov;6(3):331-43 - PubMed
  23. Exp Cell Res. 1999 Aug 25;251(1):67-78 - PubMed
  24. Cytotechnology. 2013 Jan;65(1):71-81 - PubMed
  25. Infect Immun. 2004 Aug;72(8):4689-98 - PubMed
  26. Differentiation. 1985;30(2):123-9 - PubMed
  27. Virulence. 2015;6(7):704-9 - PubMed
  28. Biol Proced Online. 2004;6:55-60 - PubMed
  29. Arch Dermatol Res. 2003 Sep;295(5):190-8 - PubMed
  30. Virol J. 2014 Jul 08;11:125 - PubMed
  31. J Cell Sci. 2012 Sep 1;125(Pt 17):3923-8 - PubMed
  32. J Invest Dermatol. 1990 Oct;95(4):409-14 - PubMed
  33. J Mater Sci Mater Med. 2016 Apr;27(4):65 - PubMed
  34. J Cell Biol. 1988 Mar;106(3):761-71 - PubMed

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