J Korean Assoc Oral Maxillofac Surg.  2007 Jun;33(3):211-220.

Role of fibroblasts in organotypic cultures of immortalized human oral keratinocytes

• Cheong JK
• Yoon KH
• Kim EC

Abstract

OBJECTIVE
In organotypic culture of immortalized human oral keratinocytes (IHOK), the change of the growth and differentiation was investigated according to the fibroblast type and the involvement of mitogen-activated protein (MAP) kinase. MATERIALS & METHODS: IHOK was cultured three dimensionally with gingival fibroblast (GF), dermal fibroblast (DF) and immortalized gingival fibroblast (IGF). We characterized biologic properties of three dimensionally reconstructed IHOK by histological, immunohistochemical, and Western blot analysis. We also investigated whether MAP kinase pathway was involved in epithelial-mesenchymal interaction by Western blot analysis.
RESULTS
The best condition of three dimensionally cultured IHOK was the dermal equivalent consisting of type I collagen and IGF. IGF increased the expression of more proliferating cell nuclear antigen (PCNA), involucrin than GF and DF in response to co-culture with IHOK. Extracellularly regulated kinase (ERK) pathway was activated in organotypic co-culture with IGF.
CONCLUSION
The organotypic co-culture of IHOK with dermal equivalent consisting of type I collagen and IGF resulted in excellent morphologic and immunohistochemical characteristics and involved ERK pathway. The epithelial-mesenchymal interaction was activated according to the fibroblast type.

Keyword

Fibroblasts; Keratinocytes; Mitogen-activated protein kinase

MeSH Terms

Blotting, Western
Coculture Techniques
Collagen Type I
Fibroblasts*
Humans*
Keratinocytes*
MAP Kinase Signaling System
Phosphotransferases
Proliferating Cell Nuclear Antigen
Collagen Type I
Phosphotransferases
Proliferating Cell Nuclear Antigen