Abstract

The development of
pharmacological compounds requires highly predictive in vitro test systems.
Assays based on primary neuronal cultures obtained from animals are commonly
used although correlation between animal and human data might be weak in some
cases. As a solution to this problem, the drug discovery field is strongly
moving towards human induced pluripotent stem-cell derived (hiPSC) neurons.
Here we developed several live-cell Ca imaging screening assays on the
Hamamatsu uCell instrument based on Cellular Dynamics’ iCell Glutaneurons.
Intracellular Ca dynamics as indicator of neuronal network activity was
recorded and modulated using different pharmacological tool compounds with
known neurophysiological effects. We focussed on three classes of experiments:
compounds with seizurogenic potential from the current NeuTox panel of HESI,
mGluR modulators alone or after pharmacological treatment of the neuronal
culture, and cultivation in the presence of amyloid-beta peptid which plays a
major role in Alzheimer disease. As astrocytes are highly important for the
neuronal network activity, we were also interested in a comparison between data
obtained from a pure iCell GlutaNeuron culture and a coculture of iCell
GlutaNeuron and iCell Astrocytes. For a subset of experiments the results were
benchmarked against data obtained from rat cortex primary neuron culture. iCell
GlutaNeurons were responsive to the tested compounds concentration-dependently
and with, in parts, higher sensitivity compared to the primary cell system. The
cocultivation with iCell Astrocytes, furthermore, had a substantial impact on
the network activity after compound treatment indicating a possibly even more
physiological reaction than iCell GlutaNeuron cultivation alone.

Acknowledgement: We thank
Cellular Dynamics International for sponsoring us with the iCell Glutaneurons
and iCell Astrocytes.