Generation, expansion, and differentiation of human pluripotent stem cell (hPSC) derived neural progenitor cells (NPCs)

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7 Citations (Scopus)

Abstract

Human pluripotent stem cell (hPSC)-derived neural progenitor cells (NPCs), a multipotent cell population that is capable of near indefinite expansion and subsequent differentiation into the various cell types that comprise the central nervous system (CNS), could provide an unlimited source of cells for neural-related cell-based therapies and disease modeling. However, the use of NPCs for the study and treatment of a variety of debilitating neurological diseases requires the development of scalable and reproducible protocols for their generation, expansion, characterization, and neuronal differentiation. Here, we describe a serumfree method for the stepwise generation of NPCs from hPSCs through the sequential formation of embryoid bodies (EBs) and neuro-epithelial-like rosettes. NPCs isolated from neural rosette cultures can be homogenously expanded while maintaining high expression of pan-neural markers such as SOX1, SOX2, and Nestin. Finally, this protocol allows for the robust differentiation of NPCs into microtubule-associated protein 2 (MAP2) and β-Tubulin-III (β3T) positive neurons.

Original languageEnglish (US)
Pages (from-to)87-102
Number of pages16
JournalMethods in Molecular Biology
Volume1212
DOIs
StatePublished - 2015

Fingerprint

Pluripotent Stem Cells
Stem Cells
Embryoid Bodies
Nestin
Microtubule-Associated Proteins
Tubulin
Cell- and Tissue-Based Therapy
Central Nervous System
Neurons
Population

Keywords

  • Embryoid bodies
  • Human embryonic stem cell (hESC)
  • Human pluripotent stem cell (hPSC)
  • Neural progenitor cells (NPC)
  • Neuronal differentiation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Medicine(all)

Cite this

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abstract = "Human pluripotent stem cell (hPSC)-derived neural progenitor cells (NPCs), a multipotent cell population that is capable of near indefinite expansion and subsequent differentiation into the various cell types that comprise the central nervous system (CNS), could provide an unlimited source of cells for neural-related cell-based therapies and disease modeling. However, the use of NPCs for the study and treatment of a variety of debilitating neurological diseases requires the development of scalable and reproducible protocols for their generation, expansion, characterization, and neuronal differentiation. Here, we describe a serumfree method for the stepwise generation of NPCs from hPSCs through the sequential formation of embryoid bodies (EBs) and neuro-epithelial-like rosettes. NPCs isolated from neural rosette cultures can be homogenously expanded while maintaining high expression of pan-neural markers such as SOX1, SOX2, and Nestin. Finally, this protocol allows for the robust differentiation of NPCs into microtubule-associated protein 2 (MAP2) and β-Tubulin-III (β3T) positive neurons.",
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