Category: Automation and High-Throughput Technologies
There is a surge of interest in fluorescent measurement of intracellular calcium oscillations in neural cells in the high-throughput manner for the neurotoxicity assessment as well as for the neural disease model study. We measured Ca2+ oscillations in rat primary and human iPSC-derived neurons to detect drug-induced neural activity changes using a kinetic plate reader FDSS that measures fluorescent signals of all wells in a plate simultaneously.
Rat cortical primary neurons were seeded and cultured in the various culture conditions (different medium and with/without nanofiber) in the 384-well plates for 14 days. Then, the cells were labeled with a calcium-sensitive fluorescent dye Cal-520 and intracellular Ca2+ oscillations were measured for 5 min with FDSS. First, we evaluated various culture conditions for producing optimal Ca2+ oscillations. Then, a set of fifteen reference compounds for excitatory and inhibitory neural activity, in particular risk compounds on seizure, were applied to the cells. The dose-dependent increase and decrease of the number of Ca2+ oscillation for 5 min (burst rate) and of the oscillation amplitudes were recorded, which are almost consistent with the reported results in rat cortical neurons and in clinical.
We also measured Ca2+ oscillations in human iPSC-derived mixed cortex neurons labeled with Cal-520 in 384-well plates and detected drug-induced synchronization of Ca2+ oscillations.
These results show that Ca2+ oscillation measurements in primary and human iPSC-derived neurons in 384-well plates on FDSS would give useful information on drug-induced neural toxicity assessment.
Natsumi Kato– Application Engineer, Hamamatsu Photonics K.K., Hamamatsu City, Shizuoka, Japan
Hamamatsu Photonics K.K.
Hamamatsu City, Shizuoka, Japan
Natsumi Kato is an application engineer at Hamamatsu in Japan since 2011. She studied Molecular Biology at the Nagoya University before joining Hamamatsu, and obtained her PhD degree in Photonics Industry Management from the Graduate School for the Creation of New Photonics Industries in 2016.