It is the purpose of this project to analyze and develop new and difficult systems for cell culture. We are now pursuing intensively a single cell system: culture of cells from the neonatal rat olfactory epithelium (OLFE). Using complex media and substrates we have succeeded in culturing several cell types from the OLFE. The mixed, mass cultures of these cells provide an appropriate conditioned medium that has permitted the isolation of >20 clonal cell strains from 3rd to 6th passage cultures. We have shown that several of our cloned cell lines have sensitive (submicromolar) and selective (different response patterns in each line) odorant-dependent second messenger responses (both cAMP and Ca4++). This fact, coupled with our demonstration that these same cell strains are positive for neuron-specific enolase, CAP43, as well as carnosine and carnosine synthetase, now establish that we have right cell type in culture. Development of this system would make available the first mammalian neuroblast to neuron cell culture system and provide a means to study the growth and differentiation dichotomy common to all blast cell systems. It is hoped that basic issues in olfactory sensory physiology can be explored with this system.
