The olfactory system can detect many odorants with high sensitivity and selectivity based on the expression of nearly a thousand types of olfactory receptors (ORs) in olfactory receptor neurons (ORNs). These ORs have a dynamic odorant detection range and contribute to signal encoding processes in the olfactory bulb (OB). To harness the capabilities of the olfactory system and develop a biomimetic sensor, stable culture and maintenance of ORNs are required. However, in vitro monolayer culture models have several key limitations: i) short available period of cultured neurons, ii) low cultural efficiency, and iii) long-term storage challenges. This study aims to develop a technique: i) to support the spheroid culture of primary ORN precursors facilitating stable maintenance and long-term storage, and ii) to demonstrate the viability of ORN spheroid culture in 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 by monitoring intracellular calcium concentration upon odorant mixture stimulation; response upon odorant stimulation were observed at least for ten days in these cultivated ORNs differentiated 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 process is a promising technique for the development of a bioelectronic nose and high-throughput odorant screening device.
Biomimetics ; Calcium ; Discrimination (Psychology) ; Electrodes ; In Vitro Techniques ; Mass Screening ; Neurons ; Nose* ; Odors ; Olfactory Bulb ; Olfactory Receptor Neurons*

Although we have multiple senses, multimedia mainly targets vision and olfaction. To expand the senses impacted by multimedia, olfactory stimulation has been used to enhance the sense of reality. Odors are primarily matched with objects in scenes. However, it is impractical to select all odors that match all objects in a scene and offer them to viewers. As an alternative, offering a single odor in a category as representative of other odors belonging to that category has been suggested. However, it is unclear whether viewers’ responses to videos with multiple odors (e.g., rose, lavender, and lily) from a category (e.g., flowers) are comparable. Therefore, we studied whether odors belonging to a given category could be similar in behavioral congruency and in the five frequency bands (delta, theta, alpha, beta, and gamma) of electroencephalogram (EEG) data collected while viewers watched videos. We conducted questionnaires and EEG experiments to understand the effects of similar odors belonging to categories. Our results showed that similar odors in a specific odor category were more congruent with videos than those in different odor categories. In our EEG data, the delta and theta bands were mainly clustered when odors were offered to viewers in similar categories. The theta band is known to be primarily related to the neural signals of odor information. Our studies showed that choosing odors based on odor categories in multimedia can be feasible.

Although we have multiple senses, multimedia mainly targets vision and olfaction. To expand the senses impacted by multimedia, olfactory stimulation has been used to enhance the sense of reality. Odors are primarily matched with objects in scenes. However, it is impractical to select all odors that match all objects in a scene and offer them to viewers. As an alternative, offering a single odor in a category as representative of other odors belonging to that category has been suggested. However, it is unclear whether viewers’ responses to videos with multiple odors (e.g., rose, lavender, and lily) from a category (e.g., flowers) are comparable. Therefore, we studied whether odors belonging to a given category could be similar in behavioral congruency and in the five frequency bands (delta, theta, alpha, beta, and gamma) of electroencephalogram (EEG) data collected while viewers watched videos. We conducted questionnaires and EEG experiments to understand the effects of similar odors belonging to categories. Our results showed that similar odors in a specific odor category were more congruent with videos than those in different odor categories. In our EEG data, the delta and theta bands were mainly clustered when odors were offered to viewers in similar categories. The theta band is known to be primarily related to the neural signals of odor information. Our studies showed that choosing odors based on odor categories in multimedia can be feasible.

Although we have multiple senses, multimedia mainly targets vision and olfaction. To expand the senses impacted by multimedia, olfactory stimulation has been used to enhance the sense of reality. Odors are primarily matched with objects in scenes. However, it is impractical to select all odors that match all objects in a scene and offer them to viewers. As an alternative, offering a single odor in a category as representative of other odors belonging to that category has been suggested. However, it is unclear whether viewers’ responses to videos with multiple odors (e.g., rose, lavender, and lily) from a category (e.g., flowers) are comparable. Therefore, we studied whether odors belonging to a given category could be similar in behavioral congruency and in the five frequency bands (delta, theta, alpha, beta, and gamma) of electroencephalogram (EEG) data collected while viewers watched videos. We conducted questionnaires and EEG experiments to understand the effects of similar odors belonging to categories. Our results showed that similar odors in a specific odor category were more congruent with videos than those in different odor categories. In our EEG data, the delta and theta bands were mainly clustered when odors were offered to viewers in similar categories. The theta band is known to be primarily related to the neural signals of odor information. Our studies showed that choosing odors based on odor categories in multimedia can be feasible.

Although we have multiple senses, multimedia mainly targets vision and olfaction. To expand the senses impacted by multimedia, olfactory stimulation has been used to enhance the sense of reality. Odors are primarily matched with objects in scenes. However, it is impractical to select all odors that match all objects in a scene and offer them to viewers. As an alternative, offering a single odor in a category as representative of other odors belonging to that category has been suggested. However, it is unclear whether viewers’ responses to videos with multiple odors (e.g., rose, lavender, and lily) from a category (e.g., flowers) are comparable. Therefore, we studied whether odors belonging to a given category could be similar in behavioral congruency and in the five frequency bands (delta, theta, alpha, beta, and gamma) of electroencephalogram (EEG) data collected while viewers watched videos. We conducted questionnaires and EEG experiments to understand the effects of similar odors belonging to categories. Our results showed that similar odors in a specific odor category were more congruent with videos than those in different odor categories. In our EEG data, the delta and theta bands were mainly clustered when odors were offered to viewers in similar categories. The theta band is known to be primarily related to the neural signals of odor information. Our studies showed that choosing odors based on odor categories in multimedia can be feasible.