Primary Culture of the Human Olfactory Neuroepithelium
The central cell type involved in the initial perception of odors and transduction of the sensory signal are the olfactory receptor neurons (ORNs) located in the olfactory neuroepithelium of the nasal cavities. The olfactory epithelium is a unique system similar to the neuroepithelium of the embryonic neural tube, in which new neurons are continually generated throughout adult life. Olfactory neurons are derived from precursor cells that lie adjacent to the basal lamina of the olfactory epithelium; these precursor cells divide several times and their progeny differentiate into mature sensory neurons throughout life. Thus, the human olfactory epithelium has the potential to be used as a tool to examine certain human disorders resulting from abnormal development of the nervous system. This chapter presents methods for primary culture of human ORNs, which have been used successfully by multiple investigators. The protocol provides a consistent, heterogeneous cell population, which demonstrates functional responses to odorant mixtures and exhibits a complex neuronal phenotype, encompassing receptors and signaling pathways pertinent to both olfaction and other aspects of CNS function. These cultured neural cells exhibit neurotransmitter pathways important in a number of neuropsychiatric disorders, and the ability to culture cells from living human subjects provides a tool for assessing cellular neuropathology at the individual patient level.