Modeling and Systemization of Cultured Cell for High Through-Put Analysis (HTA)

Abstract

In order to have practical tactics on neural drug high through-put analysis (HTA), the authors have created a new bioassay method for determining the functions of neural ion-gated channel receptors. The synapse model cell is prepared a novel cellular biosensing concept. Cloned ligand-gated ion channel receptors are integrated into insect cells, because they are easier to cultivate than mammalian cells. The engineered insect cell can express ligand-gated glutamate receptor (iGluR) on its cell surface as similar as on a post-synapse membrane. The ligand-gated ion channel receptors on the insect cell can perform functions similar to those performed on the native neural synapse membrane. We have used outer cell potential measurement to record agonist-dependent cellular responses in model cell to express ligand-gated ion channel receptor on microelectrode surface. The iGluR lets Na⁺ ions transferred into the cell, and the amount of positively charged ions on the electrode surface where the cell is attached decreases transiently. The temporary scarcity of positively charged ions causes a potential shift between the recording electrode and the reference electrode. Because the potential profile reflects the channel gate function very well, it provides information on the efficacy of transmitter analogs, information which can then be used for designing or assessing the efficacy of clinical drugs. This synapse model cell system is a good example of the development of advanced cellular biosensing techniques.