Primary Culture of Chinchilla Middle Ear Epithelium

Abstract

Chinchilla middle ear epithelium was successfully cultured in medium containing Dulbecco's modified Eagle's medium, Ham's F12 mixture, and fetal bovine serum. After 3 to 5 days, the explants produced outgrowths of primarily flat polygonal and ciliated cells that persisted for up to 10 days in culture. These cells in the outgrowth often formed a “dome” indicating the presence of functional polarization and fluid transportation capability. The ciliated cells were more frequently found near the explant, and were fewer in number in the area distant from the explant. This finding suggests that the ciliated cells in the outgrowth are migrated ciliated cells deriving from the explant. That secretory cells were not identified in the outgrowth indicated that the present culture technique did not support secretory activity. Using the present culture technique, we were able to maintain the explants and primary cultured cells for up to 14 days in a majority of cases; hence, these techniques appear to be applicable to a number of in vitro studies.