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Curr Protoc Stem Cell Biol. 2017 Aug 14;42:2D.20.1-2D.20.14. doi: 10.1002/cpsc.31.

Generation of Oligodendrogenic Spinal Neural Progenitor Cells From Human Induced Pluripotent Stem Cells.

Khazaei M1, Ahuja CS1,2,3, Fehlings MG1,2,3,4,5.

Author information

1
Division of Genetics and Development, Krembil Research Institute, Toronto, Ontario, Canada.
2
Institute of Medical Science, University of Toronto, Toronto, Ontario, Canada.
3
Division of Neurosurgery, University of Toronto, Toronto, Ontario, Canada.
4
Spinal Program, Toronto Western Hospital, University Health Network, Toronto, Ontario, Canada.
5
Faculty of Medicine, University of Toronto, Toronto, Ontario, Canada.

Abstract

This unit describes protocols for the efficient generation of oligodendrogenic neural progenitor cells (o-NPCs) from human induced pluripotent stem cells (hiPSCs). Specifically, detailed methods are provided for the maintenance and differentiation of hiPSCs, human induced pluripotent stem cell-derived neural progenitor cells (hiPS-NPCs), and human induced pluripotent stem cell-oligodendrogenic neural progenitor cells (hiPSC-o-NPCs) with the final products being suitable for in vitro experimentation or in vivo transplantation. Throughout, cell exposure to growth factors and patterning morphogens has been optimized for both concentration and timing, based on the literature and empirical experience, resulting in a robust and highly efficient protocol. Using this derivation procedure, it is possible to obtain millions of oligodendrogenic-NPCs within 40 days of initial cell plating which is substantially shorter than other protocols for similar cell types. This protocol has also been optimized to use translationally relevant human iPSCs as the parent cell line. The resultant cells have been extensively characterized both in vitro and in vivo and express key markers of an oligodendrogenic lineage.

KEYWORDS:

differentiation; human induced pluripotent stem cell; neural progenitor; oligodendrocyte; spine

PMID:
28806852
DOI:
10.1002/cpsc.31
[Indexed for MEDLINE]

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