High-throughput drug screening (HTS) campaigns are commonly performed using immortalized recombinant cell lines that overexpress the target of interest. But for some complex targets, such as NMDAR, this approach could be very challenging, since it is a multi-subunit channel and, furthermore, its overexpression in non-neuronal cells results in cell death due to its constitutive activation at depolarized membrane potential.
The development of high-quality neuronal cultures derived from human induced pluripotent stem cells (iPSCs), such as iCell® GlutaNeurons (FCDI), for compound screening, offers many advantages over recombinant cell lines, avoiding the issues related to the non-physiological overexpression of the target and allowing to use relevant cellular disease models.
Here we report the development of a cell-based assay, in miniaturized 384-well plate format, using the iCell® GlutaNeurons, to study the modulation of NMDARs and other different calcium players, having a pivotal role in different neuro-disorders.
A pilot screening using the LOPAC®1280 library (Library of Pharmacologically Active Compounds), which contains 1280 well-characterized compounds with annotated activity, was performed and intracellular calcium variations were monitored.
The very promising obtained results demonstrate the feasibility to use iCell® GlutaNeurons, in HTS campaigns, to identify small molecule agonists, antagonists and modulators for NMDARs, and other relevant neuronal targets, inducing intracellular calcium waves.
stem cells, hiPSC cells, glutamatergic neurons, cell-based assays, NMDAR and glutamatergic receptors, MTS/HTS and screening campaigns, calcium mobilization assay