Long-term natural culture of cochlear sensory epithelia of guinea pigs

To culture and maintain mammalian cochlear cells in vitro is still a big challenge. Only immortalized cochlear cell lines are available. With refinement of culture media and techniques, cochlear sensory epithelial cells of guinea pigs have been cultured without any genetic manipulation using a modified Keratinocyte medium for more than 6 months. The isolated cell clones by cloning cylinders showed a large, flat, epithelial morphotype and expressed cytokeratin and a tight junction associated protein ZO-1, but did not express vimentin. These cells were also labeled with Brn3.1 and calretinin, which are regarded as early hair cell markers. The immunostaining confirmed the culture cells derived from cochlear sensory epithelia. These non-immortalized natural cochlear cells provide valuable cell sources for molecular and genetic studies of the inner ear.