Many technical challenges exist in the co-culture of multiple types of cells, including medium optimization, cell-to-cell connection, and selective data acquisition of cellular responses. Particularly, mixed cellular responses limit the precise interpretation of intercellular signal transduction. Here, we report the formation of an agarose gel skin on neurons closely assembled with gustatory cells to selectively stimulate gustatory cells by retarding the diffusion of tastants to neurons. The signal transmission, triggered by denatonium benzoate, from gustatory cells to neurons was monitored using intracellular calcium ion concentrations. The agarose gel skin efficiently suppressed the direct transfer of tastants to neurons, decreasing the number of responsive neurons from 56 to 13% and the number of calcium ion signals per neuron from multiple to single. The assembly of neurons with gustatory cells induced the high level of neuronal responses through taste signal transduction from gustatory cells to neurons. However, the calcium ion signal peaks of free neurons coated with agarose gel were much shorter and weaker than those of neurons closely assembled with gustatory cells. This work demonstrated that agarose gel skin is a simple, fast, and effective means to increase the signal selectivity of cellular responses in the co-culture of multiple types of cells.