Immortalization of Chinchilla Middle Ear Epithelial Cells by Adenovirus 12-Simian Virus 40 Hybrid Virus

Abstract

In order to study the cellular and molecular mechanisms of the pathogenesis of otitis media, a chinchilla middle ear epithelial cell line (CMEE-1) with differentiated cell characteristics was established by infection of a primary culture with the adenovirus 12-simian virus 40 (Ad12-SV40) hybrid. This cell line has been in continuous culture for 42 passages, whereas the parent cells underwent senescence and died at the 8th passage. The cell line also retains epithelial morphology and expresses cytokeratin polypeptides 4, 7, and 18, characteristic markers for epithelia. In Western blots of cell proteins, bands at 94 and 53 kd were labeled after binding antibodies against SV40 large T antigen and p53, respectively. Karyotype analysis showed that the cell line is derived from chinchilla epithelial cells. These findings confirm that the cell line is a chinchilla epithelial cell immortalized by the hybrid virus.

Influenza A Virus Infection of Human Middle Ear Cells In Vitro

“…Our experience was similar to that described by Nakamura et al 15 using chinchilla‐derived middle ear mucosal cells, as well as by other investigators. 16–18 In the present study, cells retained morphological and immunocytochemical properties of epithelial cells for nearly 2 months. Prolonged passage of these cells ultimately resulted in a change to a spindle morphology coincident with a loss of cytokeratin staining although cytokeratin 18 mRNA was still detectable by RT‐PCR.…”

In vitroreconstruction of a three-dimensional middle ear mucosal organ and itsin vivotransplantation

“…The culture of middle ear mucosal epithelial cells is difficult in comparison with the culture of mucosal epithelial cells from the skin or the paranasal cavity mucosa. There have been various reports regarding the culture of middle ear mucosal epithelial cells from humans [14Á19], rats [20], chinchillas [21], etc. However, unlike the case for skin keratinocytes, there is still no consensus on the method for their culture.…”

Cell lines in inner ear research

“…They are sealed by well-developed tight junctions and are able to form epithelial domes. The Chinchilla middle ear epithelial cell line, CMEE-1, was established by infecting a primary culture with an adenovirus 12-SV40 virus hybrid (Jin et al, 1999). This cell line retains epithelial features and expresses different cytokeratins.…”