iPSC Culturing
Maintenance, Expansion, Selection, and Differentiation
My iPSC culturing, maintenance, and differentiation training was done in association with the EDDU, whose protocols generate some of the most homogenous cultures in the world. In our latest project, iPSCs from a total of 9 different donors were used for modeling Parkinson's disease pathology in vitro.
iPSC Characterization
Antibody profiling and Electrical Activity
Characterization
Following iPSC differentiation, neuronal maturation represents the most time-intensive stage of the culture process, typically requiring 2–6 weeks. To achieve full dopaminergic (DA) identity—marked by the expression of DAT and GIRK2—at least 4 weeks of maturation are necessary. A similar timeline applies to cortical neurons, which require a minimum of 4 weeks, and ideally 6, to reach maturity and exhibit both VGLUT1 and GABA expression.
Electrical Activity
Another approach to confirm neuronal identity, following molecular and morphological characterization, is to examine their electrophysiological properties. To this end, we plated neurons onto microelectrode arrays (MEAs), which enable non-invasive recordings of spontaneous electrical activity. From these recordings, I generated raster plots to visualize firing patterns across different neuronal populations and extracted representative waveforms from individual electrodes to assess action potential architecture during MEA recordings.