The phenotype of in vitro reconstituted normal human oral epithelium is essentially determined by culture medium.
OBJECTIVE
To evaluate the role of various culture media and serum supplement on growth of oral cells in monolayer, and on morphogenesis of in vitro reconstituted normal human oral epithelium.


METHODS
Primary keratinocytes and fibroblasts were isolated from normal human buccal mucosa. The monolayers were assessed by growth curve analysis and morphology. The organotypic cultures were evaluated by morphometry, immunohistochemistry, and TUNEL.


RESULTS
FAD medium (a 3:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium) was able to support fibroblast growth in defined conditions, and to diminish the negative effect of physiological Ca concentration on keratinocytes in monolayers. Medium type had a profound influence on morphogenesis of in vitro reconstituted human oral epithelium. FAD medium was superior to other types of medium tested in supporting both epithelial growth and differentiation. Defined conditions supported epithelial morphogenesis equally well as serum-containing medium.


CONCLUSIONS
This study points to an essential role of medium composition for optimized growth and differentiation of primary organotypic cultures.
Costea
Daniela Elena
DE
Department of Odontology, Oral Pathology and Forensic Odontology, Faculty of Dentistry, University of Bergen, N-5021 Bergen, Norway.
Dimba
Anne Okumo Elizabeth
AO
Loro
Lado Lako
LL
Vintermyr
Olav Karsten
OK
Johannessen
Anne Christine
AC
D001769:Blood; D016923:Cell Death; D002454:Cell Differentiation; D002478:Cells, Cultured; D003470:Culture Media / Q000145:classification; D004848:Epithelium / Q000254:growth & development / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D015603:Keratinocytes / Q000378:metabolism; D009024:Morphogenesis; D009061:Mouth Mucosa / Q000254:growth & development / Q000378:metabolism*; D010641:Phenotype; D046509:Tissue Culture Techniques
D003470:Culture Media