Spheroid culture of rat olfactory receptor neurons and its application for bioelectronic nose

Abstract

A novel concept of cell-based biosensor mimicking odorants encoding process of the mammalian ORN was proposed and developed. Rat ORN precursor spheroids were formed by using recombinant protein (REP; TGPG[VGRGD(VGVPG)6]20WPC) and used as a bioelement and combined with commercialized multi electrode array (MEA). Spheroid culture of ORN precursors realized stable maintenance and long-term storage of ORN precursors with preservation of stemness characters. Physiological characters of the differentiated ORNs from spheroids were verified by monitoring change of intracellular calcium concentration upon odorant mixture stimulation and these characters were well preserved in the long-term cultured ORN spheroids. Lastly, differentiated ORN on the MEA generated electrical signals upon odorants stimulation and these signals were collected as signal patterns and analyzed for detecting and discriminating odorant types and concentrations. Developed ORN-based biosensor showed superior repeatability and reproducibility in the odorant detection and the ORN spheroid culture have potentials to be applied in the development of a bioelectronic nose and high-throughput odorant screening.