A Guide to Generating and Using hiPSC Derived NPCs for the Study of Neurological Diseases

Abstract

Post-mortem studies of neurological diseases are not ideal for identifying the underlying causes of disease initiation, as many diseases include a long period of disease progression prior to the onset of symptoms. Because fibroblasts from patients and healthy controls can be efficiently reprogrammed into human induced pluripotent stem cells (hiPSCs), and subsequently differentiated into neural progenitor cells (NPCs) and neurons for the study of these diseases, it is now possible to recapitulate the developmental events that occurred prior to symptom onset in patients. We present a method by which to efficiently differentiate hiPSCs into NPCs, which in addition to being capable of further differentiation into functional neurons, can also be robustly passaged, freeze-thawed or transitioned to grow as neurospheres, enabling rapid genetic screening to identify the molecular factors that impact cellular phenotypes including replication, migration, oxidative stress and/or apoptosis. Patient derived hiPSC NPCs are a unique platform, ideally suited for the empirical testing of the cellular or molecular consequences of manipulating gene expression. Video LinkThe video component of this article can be found at

Gene expression elucidates functional impact of polygenic risk for schizophrenia

“…hiPSC forebrain NPCs were derived from the three controls as described previously 74 . These samples were selected irrespective of their genotypes for the FURIN -eQTL SCZ-risk variant at SNP rs4702, with two being heterozygous G/A and the third homozygous risk G/G.…”

THC Treatment Alters Glutamate Receptor Gene Expression in Human Stem Cell-Derived Neurons

“…Passage-matched hiPSCs were used for all experiments. Cohort 1 NPCs were derived from hiPSCs using dual-SMAD inhibition as previously described [39,40]. NPCs were cultured on Matrigel (BD)-coated plates in NPC media (DMEM/F12, 1× N2 [Life Technologies], 1× B27-RA [Life Technologies], and 20 ng/mL FGF2), maintained at high density and passaged with Accutase (Millipore).…”

Genetic basis of neurological disorder with human induced pluripotent stem cells

“…Recent discovery in the field of stem cell technology accessed the researchers to produce hiPSCs (Yu et al, 2007). Today hiPSCs model are used to understand the molecular mechanism and features of neurological disease (Aaron Topol et al, 2015).…”