Establishment and Characterization of an In Vitro Model for Cholesteatoma

다국어 초록 (Multilingual Abstract)

Objectives. Experimental models are of importance to study the pathogenesis of middle ear cholesteatoma, however, they
were not established until now. We aimed to develop in vitro model of middle ear cholesteatoma using primary
keratinocytes and fibroblasts isolated from cholesteatoma tissue. HaCaT cell line was used as a “skin equivalent”
and to compare the grade of homogeneity between cholesteatoma keratinocytes and HaCaT cells.
Methods. Primary keratinocytes were isolated from cholesteatoma tissue, co-cultured with preliminary prepared feeder
layer from cholesteatoma fibroblasts and subsequently air-exposed. The protein profile of cholesteatoma keratinocytes
and HaCaT cells was evaluated by means of immunoblot using monoclonal antibody against cytokeratin (CK) 13
and 16. Tissue localization of CK 13 and 16 was accomplished with immunohistochemistry.
Results. Different protein profile and stronger expression of CK 13 and 16 were demonstrated in cholesteatoma keratinocytes
in comparison with HaCaT cells. Bigger stratification was observed in the 3D-in vitro systems when both cholesteatoma
keratinocytes and HaCaT cells were respectively co-cultured with fibroblasts in comparison with the corresponding
control groups without fibroblasts.
Conclusion. 3D-model demonstrates the significance of intercellular interaction between components of cholesteatoma
tissue.

다국어 초록 (Multilingual Abstract)

Objectives. Experimental models are of importance to study the pathogenesis of middle ear cholesteatoma, however, they
were not established until now. We aimed to develop in vitro model of middle ear cholesteatoma using primary
keratinocytes and fibroblasts isolated from cholesteatoma tissue. HaCaT cell line was used as a “skin equivalent”
and to compare the grade of homogeneity between cholesteatoma keratinocytes and HaCaT cells.
Methods. Primary keratinocytes were isolated from cholesteatoma tissue, co-cultured with preliminary prepared feeder
layer from cholesteatoma fibroblasts and subsequently air-exposed. The protein profile of cholesteatoma keratinocytes
and HaCaT cells was evaluated by means of immunoblot using monoclonal antibody against cytokeratin (CK) 13
and 16. Tissue localization of CK 13 and 16 was accomplished with immunohistochemistry.
Results. Different protein profile and stronger expression of CK 13 and 16 were demonstrated in cholesteatoma keratinocytes
in comparison with HaCaT cells. Bigger stratification was observed in the 3D-in vitro systems when both cholesteatoma
keratinocytes and HaCaT cells were respectively co-cultured with fibroblasts in comparison with the corresponding
control groups without fibroblasts.
Conclusion. 3D-model demonstrates the significance of intercellular interaction between components of cholesteatoma
tissue.

참고문헌 (Reference)

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