High-throughput (Hamamatsu FDSS/μCell) Live Cell Imaging Services
Our higher-throughput working horse with parallel recordings in 96- or 384-well format at a sampling frequency of up to 120 Hz.
Intracellular Ca2+ Imaging of spontaneously beating cardiomyocytes using the higher throughput Hamamatsu FDSS/μCell system. A) Positive inotropic effect. Application of this beta2-sympathomimetic compound results in a concentration-dependent increase of intracellular Ca2+ concentration during each beat. B) A Ca2+ channel inhibitor both reduces Ca2+ transient amplitude and duration, resulting in negative inotropicity. C) Application of a potent hERG channel inhibitor results in the detection of proarrhythmic behavior in the investigated cardiomyocytes.
Hamamatsu user meeting June 2018 in Basel
Bringing human neuronal biology to HTS: Functional Drug Screening with iCell GlutaNeurons and Astrocyte on the Hamamatsu FDSS/µCELL.
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 the 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 μCell instrument based on Cellular Dynamics’ iCell Glutaneurons. Intracellular Ca dynamics as an 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 peptide 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.
Acknowledgment: We thank Cellular Dynamics International for sponsoring us with the iCell Glutaneurons and iCell Astrocytes.
Loser, D. et. al. Acute effects of the imidacloprid metabolite desnitro-imidacloprid on human nACh receptors relevant for neuronal signaling. Molecular Toxicology 2021 | Contact an author.
Loser, D et. al. Functional alterations by a subgroup of neonicotinoid pesticides in human dopaminergic neurons. In vitro systems 2021 | Contact an author.
Loser, D et. al. Human neuronal signaling and communication assays to assess functional neurotoxicity. Organ Toxicity and Mechanisms 2021 | Contact an author.