An original organotypic culture method to study the organ of Corti of the newborn rat in vitro

Abstract

The inner ear growth is related to several factors which are not yet well known. In order to study the effect of growth factors on the development of the auditory receptor cells, we have chosen first to establish an in vitro model of the inner ear. Newborn rats were selected as the source of tissue because the inner ear is immature enough at this stage of development for studying some relations between receptors and their innervation. Until recently, the Maximov slide assembly technique was the only organ culture system available and silver neurofibrillary methods were used to stain the nervous structures. These are difficult and time-consuming techniques. With the use of a collagen gel drop floating in the culture medium, we have developed a simple and reliable method. Furthermore, an immunohistochemical fiber-staining technique with anti-neurofilament and histochemical staining technique with phalloidin allows us to check in a few days the organotypy at the spiral ganglion and hair cell levels. This floating drop method gave us some preliminary information about the spiral neuron cells which survived.