Comparison and Optimization of hiPSC Forebrain Cortical Differentiation Protocols
Name
Muratore-2014-Comparison and Optim.pdf
Size
23.33 MB
Format
Adobe PDF
Checksum (MD5)
d8bbb13f48b50c401e3a13a8731a8614
Author(s)
Muratore, Christina R.
Srikanth, Priya
Callahan, Dana G.
Young-Pearse, Tracy L.
Date Issued
August 2014
Journal
PLoS ONE
Publisher
Public Library of Science
Citation
Muratore, Christina R., Priya Srikanth, Dana G. Callahan, and Tracy L. Young-Pearse. “Comparison and Optimization of hiPSC Forebrain Cortical Differentiation Protocols.” Edited by Mirella Dottori. PLoS ONE 9, no. 8 (August 28, 2014): e105807.
Version
Final published version
Abstract
Several protocols have been developed for human induced pluripotent stem cell neuronal differentiation. We compare several methods for forebrain cortical neuronal differentiation by assessing cell morphology, immunostaining and gene expression. We evaluate embryoid aggregate vs. monolayer with dual SMAD inhibition differentiation protocols, manual vs. AggreWell aggregate formation, plating substrates, neural progenitor cell (NPC) isolation methods, NPC maintenance and expansion, and astrocyte co-culture. The embryoid aggregate protocol, using a Matrigel substrate, consistently generates a high yield and purity of neurons. NPC isolation by manual selection, enzymatic rosette selection, or FACS all are efficient, but exhibit some differences in resulting cell populations. Expansion of NPCs as neural aggregates yields higher cell purity than expansion in a monolayer. Finally, co-culture of iPSC-derived neurons with astrocytes increases neuronal maturity by day 40. This study directly compares commonly employed methods for neuronal differentiation of iPSCs, and can be used as a resource for choosing between various differentiation protocols.
MIT Department
Harvard University--MIT Division of Health Sciences and Technology
Terms of Use
Creative Commons Attribution
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1371/journal.pone.0105807
