ISSCR – International Society for Stem Cell Research
Essen Booth #736
June 14th-17th — Boston, MA, USA
Boston Convention and Exhibition Center (BCEC)
Visit our booth at ISSCR to learn about the NEW Next-Generation IncuCyte® S3 Live-Cell Analysis System and the latest trends and applications in live-cell analysis.
From the moment cells are placed in culture, the learning opportunity begins. With the IncuCyte S3 Live-Cell Analysis System you can continuously monitor stem cell reprogramming and differentiation events – right inside your incubator. The IncuCyte automatically acquires and analyzes images around the clock, for days, weeks, even months, enabling you to derive deeper and more physiologically relevant insights into stem cell reprogramming and differentiation processes from real-time kinetic data.
Also, stop by for a chance to win prizes! Spin the wheel of fortune to win memory sticks, mugs and more.
At the show, join our poster presentation presented by Dr. Aaron Overland, Senior Application Scientist, to learn the benefits of incorporating real-time, automated cell imaging and analysis to your in vitro assay workflow for stem cell research.
Quantitative Live-cell Analysis for Optimization of Culture Conditions and Evaluation of Cell Health In Human Induced Pluripotent Stem Cell-derived Neurons
Presenter: Aaron Overland, Ph.D., Senior Application Scientist
Session Date and Time: Thursday, June 15, 2017 7:00 PM – 8:00 PM
Location: POSTER SESSION II-EVEN
Poster Board Number: T-1134
Authors: Aaron Overland1, Daniel Apledorn1, John Rauch1, Libby Oupicka1, David Rock1. 1Essen Bioscience
A major limitation in studying human diseases affecting the nervous system is the ability to culture, monitor and analyze neuronal cells that accurately represent human phenotypes of these disorders. The use of human induced pluripotent stem cell (hiPSC)-derived neurons has provided a new approach aimed at modeling neurological diseases. Monitoring neuronal morphology and cell health in long-term culture is critical for the characterization and evaluation of these novel model systems. Traditional approaches rely on endpoint assays and imaging techniques that require immunochemical staining, which provide limited real-time kinetic information. Here we present data highlighting optimal culture conditions for cell viability and neurite outgrowth in hiPSC-derived neurons from Cellular Dynamic International (CDI, iCell Neurons). We also evaluated neurite outgrowth and cellular viability in iCell Gluta Neurons from CDI using a quantitative, live-cell imaging approach with the IncuCyte ® Live-Cell Analysis System over days/weeks in culture. Testing multiple culture and plate coating conditions, we find iCell Neurons respond best with a combination of BrainPhys media (Stem Cell Technologies) and iCell DOPA Neuron Supplement/iCell Nervous System Supplement (CDI) combined with a plate coating of polyethyleneimine and laminin. To exemplify a real-time imaging approach using hiPSC-derived neurons, we evaluated glutamate- and kainate-induced excitotoxicity using the IncuCyte ® phase/fluorescent NeuroTrack applications multiplexed with Annexin V reagents in iCell Gluta Neurons. Glutamate and kainate produced a concentration- and time-dependent decrease in neurite length with a concomitant increase in red or green object count (indicating cell death) over 72 hours. Glutamate and kainate produced a similar effect when measured by IncuCyteT Cytotox Red, IncuCyteT Cytotox Green and caspase 3/7 reagents. Treatment with the NMDA receptor antagonist MK-801 and the AMPA receptor antagonist NBQX reduced the glutamate- and kainate-induced effects on neurite length and cell death. These data outline optimal culture conditions for iCell Neurons and demonstrate the ability of the IncuCyte ® approach for real-time, long-term quantitative analysis of iPSC-derived neuronal cell health.