Chapter 7. Human Oral Epithelium

  1. R. Ian Freshney and
  2. Mary G. Freshney
  1. Roland C. Grafström

Published Online: 28 APR 2002

DOI: 10.1002/0471221201.ch7

Culture of Epithelial Cells, Second Edition

Culture of Epithelial Cells, Second Edition

How to Cite

Grafström, R. C. (2002) Human Oral Epithelium, in Culture of Epithelial Cells, Second Edition (eds R. I. Freshney and M. G. Freshney), John Wiley & Sons, Inc., New York, USA. doi: 10.1002/0471221201.ch7

Editor Information

  1. CRC Beatson Laboratories, Glasgow, Scotland

Author Information

  1. Experimental Carcinogenesis, Inst. Environmental Medicine, Karolinska Institutet, S-171 77 Stockholm, Sweden

Publication History

  1. Published Online: 28 APR 2002
  2. Published Print: 12 APR 2002

Book Series:

  1. Culture of Specialized Cells

Book Series Editors:

  1. R. Ian Freshney

Series Editor Information

  1. CRC Beatson Laboratories, Glasgow, Scotland

ISBN Information

Print ISBN: 9780471401216

Online ISBN: 9780471221203



  • oral epithelium;
  • keratinocytes;
  • epithelial cell culture;
  • serum-free medium;
  • EMHA;
  • MCDB 153;
  • medium preparation;
  • organotypic culture;
  • colony forming efficiency;
  • cytotoxicity;
  • carcinogenesis


After a brief introduction of the epithelial structures found in the oral mucosa, a review of the methods utilized by various investigators for culture of nonmalignant oral epithelium is presented including a tabulated presentation of the respective research areas and results. Technical aspects applicable to monolayer, multilayer, explant, and organotypic culture are summarized. Subsequently, detailed protocols for serum-free culture of oral epithelium are shown based on the experiences derived from specimens obtained from more than 800 individuals over the last two decades. Step-by-step protocols for media fabrication include information on commercial source, preparation, and storage for each of the components. The basic protocols for deriving, handling, and storage of cells include primary and transfer culture at low (clonal) and high density. The overall information presented demonstrates that basic laboratory resources are sufficient to reproducibly generate reagents and conditions for oral keratinocyte culture from single chemicals and bovine pituitaries without the necessity of purchasing buffers and media from commercial sources. Notably, the conditions developed for normal oral keratinocytes are also applicable to at least some immortalized (nonmalignant) and malignant variants in both monolayer and organotypic culture.