Olfactory Receptor Neurons in Partially Purified Epithelial Cell Cultures:Comparison of Techiques for Partial Purification and Identification of Insulin as an Important Survival Factor

摘要

Olfactory neurons have the rare property of being replaceo :nrc~.gb~out fe Pactors regulating different developmental stages of olfactory receptor neurons (ORNs) are of great interesi, beca-~se s..~ factors might be used to extend regeneration in the post-developmental brain and spinal cord. Also, cnese factocs ma~ potentiaily be exploited to treat various smell disorders arising from changes in the olfactory epitheliurri CnaracteinzatOfl of trophic factors for ORNs requires cell culture systems that are simple and easy to manipulate. We have compared f·c~ur different cell culture preparations, using two different enzymes and two different media to develop a simple cJture s~,5tern of olfactory epithelial cells. Our preferred preparation, which produces partially purified olfactory epithehal cjcures, oses :r’~’psin oFssociation and a serum-free keratinocyte growth medium (KGM) supplemented with insulin. These conc tiers Siupport OR survival up to 1 week. They also supported other elements of the olfactory epithelium such as Bowmars gland c~lls and horizontal basal cells. Olfactory epithelial cells predominate, while contaminating mesenchymal cells Igla and ~brobastsI) are present in low numbers. Using these cultures, it was determined that insulin was required for ORN sur’~ ~aI n vitro The simplicity of the epithelial cultures will be useful for further studies of insulin and other ORN tropnic factors.