Longitudinal Characterization of Human Stem Cell-Derived Neurons Using Calcium Imaging
The ability to measure neuronal activity and network activity in a longitudinal manner is paramount to the study of neurological disease Due to the synaptic plasticity of neuronal cell cultures, identifying dynamic changes between normal and mutated models is critical for understanding changes in network connectivity. To evaluate activity differences between normal and diseased states, we used mutant and control human induced pluripotent stem cells (iPSCs) to generate neuronal cultures by forcing expression of Neurogenin-2. Using the IncuCyte® S3 live-cell analysis system, we measured neuronal activity over the course of several weeks for functional analysis of differences in activity and connectivity. We found that iPSC-derived neurons had increased activity over time and formed synchronous neuronal networks around 3 weeks of differentiation. Data generated showed a reproducible, relevant phenotypic readout for long-term network changes in this epileptic cell model.
Key webinar discussion topics include:
- Differentiate iPSCs into neurons using forced Neurogenin-2 expression
- Acquire and analyze calcium imaging data over time, using the IncuCyte® S3 live-cell analysis system
- The importance of long-term analysis of neuronal activity to evaluate differences in activity and connectivity for iPSC derived neurons
About the speaker
Louis Dang, MD/PhD
University of Michigan
Louis Dang is a pediatric epileptologist and neuroscientist at the University of Michigan. His research focuses on how genetic mutations cause early-life epilepsies. Using patient-derived and gene-edited stem cells, he generates 2D and 3D cultures to determine the molecular pathogenesis of epilepsy, opening new avenues for targeted therapies.