Open AccessHighly Expandable Human iPS Cell–Derived Neural Progenitor Cells (NPC) and Neurons for Central Nervous System Disease Modeling and High‐Throughput Screening
Author(s) -
Cheng Chialin,
Fass Daniel M.,
FolzDonahue Kat,
MacDonald Marcy E.,
Haggarty Stephen J.
Publication year - 2017
Publication title -
current protocols in human genetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.282
H-Index - 30
eISSN - 1934-8258
pISSN - 1934-8266
DOI - 10.1002/cphg.33
Subject(s) - induced pluripotent stem cell , reprogramming , neural stem cell , cell sorting , biology , progenitor cell , neuroscience , neural cell , cell , cell type , stem cell , microbiology and biotechnology , embryonic stem cell , genetics , gene
Abstract Reprogramming of human somatic cells into induced pluripotent stem (iPS) cells has greatly expanded the set of research tools available to investigate the molecular and cellular mechanisms underlying central nervous system (CNS) disorders. Realizing the promise of iPS cell technology for the identification of novel therapeutic targets and for high‐throughput drug screening requires implementation of methods for the large‐scale production of defined CNS cell types. Here we describe a protocol for generating stable, highly expandable, iPS cell–derived CNS neural progenitor cells (NPC) using multi‐dimensional fluorescence activated cell sorting (FACS) to purify NPC defined by cell surface markers. In addition, we describe a rapid, efficient, and reproducible method for generating excitatory cortical‐like neurons from these NPC through inducible expression of the pro‐neural transcription factor Neurogenin 2 (iNgn2‐NPC). Finally, we describe methodology for the use of iNgn2‐NPC for probing human neuroplasticity and mechanisms underlying CNS disorders using high‐content, single‐cell‐level automated microscopy assays. © 2017 by John Wiley & Sons, Inc.