The Three-Dimensional Co-Culture of Cholesteatomatous Keratinocyte and Fibroblast

Kim, Sok Chon; Hong, Nam Pyo; Choo, Jae Hak; Park, Soo Hong; Hong, Seok Chan; Cha, Chang Il

Abstract

BACKGROUND AND OBJECTIVES: There has been considerable research interest in cholesteatoma in recent years but an understanding of the pathophysiology has been handicapped by the lack of a suitable research model. Recent advances in culture methods have contributed to the study of normal physiology or pathogenesis at the level of cells. However, the conventional keratinocyte culture of cholestearoma has some weak points, since it is done under a condition that is different from cellular environments and it cannot investigate cellular interaction among keratinocytes and other cells of cholesteatoma. The purpose of this study is as follows : first, we organize a new study model for cholesteatoma using a three-dimensional (3-D) culture method; and then we study fibroblast effects on keratinocytes under the 3-D culture system.
MATERIALS AND METHODS
Aural cholesteatomas were obtained during middle ear operations. The keratinocytes and fibroblasts were first cultured separately and then co-cultured three-dimensionally. The pureness of keratinocyte layer was examined immunohistochemically. We studied fibroblast effects on keratinocytes under the 3-D culture system from the viewpoint of proliferation and differentiation.
RESULTS
We could establish 3-D culture of keratinocytes under the influence of fibroblasts. The keratinocytes of the co-culture system showed more proliferation and less differentiation than keratinocytes under the same conditions except for anchoring fibroblasts.
CONCLUSION
In this study, we successfully organized the 3-D culture system, a new study model for cholesteatoma, and identified fibroblast effects on keratinocytes. This new study model might be an effective tool for research on the interactions among various cells of cholesteatoma: however, further investigation and development would he necessary.