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Spheroid Culture of Mammalian Olfactory Receptor Neurons: Potential Applications for a Bioelectronic Nose
- Spheroid Culture of Mammalian Olfactory Receptor Neurons: Potential Applications for a Bioelectronic Nose
- Kim, Sam Hwan; Kim, So Yeun; Choi Seong-Kyun; Bae, Jisub; Jeon, Won Bae; Jang, Jae Eun; Moon, Cheil
- DGIST Authors
- Choi Seong-Kyun; Jeon, Won Bae; Jang, Jae Eun; Moon, Cheil
- Issue Date
- Experimental Neurobiology, 26(7), 574-598
- Article Type
- Author Keyword
- Cell-based biosensor; Recombinant protein; Olfactory receptor neuron; Spheroid culture; Micro electrode array
- SENSORY NEURONS; ACTIVATION; PROTEIN; DISCRIMINATION; SURVIVAL; SIGNALS; SYSTEM; SMELL
- The olfactory system can detect various odorants with high sensitivity and selectivity due to expression of nearly a thousand types of odorant receptors (ORs) in olfactory receptor neurons (ORNs). These ORs have a dynamic odorant detection range and contribute to signal encoding in the olfactory bulb (OB). To harness the capabilities of the olfactory system and develop a biomimetic sensor, stable ORN culture may be required. However, current in vitro monolayer ORN cultures have several critical limitations: short available period of cultured neurons, low cultural efficiency, and long-term storage challenges. Our study aims to develop a technique: to support the spheroid culture of primary ORN precursors facilitating stable maintenance and long-term storage, and to demonstrate the feasibility of ORN spheroid culture for developing an olfactory system mimetic bioelectronic nose. Recombinant protein (REP; TGPG[VGRGD(VGVPG)6]20WPC) was used to form the ORN spheroids. Spheroid formation enabled preservation of primary cultured ORNs without a significant decrease in viability or the expression of stemness markers for ten days. Physiological characteristics of the ORNs were verified monitoring intracellular calcium concentration upon odorant stimulation; we could observe calcium responses upon odorant stimulation for ten cultivation days in the differentiated ORNs from spheroids. Coupling ORNs with multi electrode array (MEA) enabled the detection and discrimination of odorants by analyzing the electrical signal patterns generated following odorant stimulation. Taken together, the ORN spheroid culture can be a highly promising technique for the development of bioelectronic noses or high-throughput odorant screening devices.
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