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Ntera2: A model system to study dopaminergic differentiation of human embryonic stem cells

Academic Article
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Overview

authors

  • Schwartz, C. M.
  • Spivak, C. E.
  • Baker, S. C.
  • McDaniel, T. K.
  • Loring, Jeanne
  • Nguyen, C.
  • Chrest, F. J.
  • Wersto, R.
  • Arenas, E.
  • Zeng, X. M.
  • Freed, W. J.
  • Rao, M. S.

publication date

  • October 2005

journal

  • Stem Cells and Development  Journal

abstract

  • NTera2, a human embryonal carcinoma (EC) stem cell line, shares many characteristics with human embryonic stem cells (hESCs). To determine whether NTera2 can serve as a useful surrogate for hESCs, we compared global gene expression between undifferentiated NTera2, multiple undifferentiated hESC cell lines, and their differentiated derivatives, and we showed that NTera2 cells share multiple markers with hESCs. Similar to hESCs, NTera2 cells differentiated into TH-positive cells that express dopaminergic markers including AADC, DAT, Nurr1, TrkB, TrkC, and GFRA1 when co-cultured with PA6 cells. Flow cytometry analysis showed that tyrosine hydroxylase (TH) and neural cell adhesion molecule (NCAM) expression increased, whereas SSEA4 expression decreased as cells differentiated. Medium conditioned by PA6 cells stimulated differentiation of NTera2 cells to generate TH-positive cells that expressed dopaminergic markers. Flow cytometry selected polysialylated (PSA-NCAM) cells responded to medium conditioned by PA6 cells by differentiating into TH-positive cells and expressed dopaminergic markers. Sorted cells differentiated for 4 weeks in PA6 cell conditioned media included functional neurons that responded to neurotransmitters and exhibited electronic excitability. Therefore, NTera2 cell dopaminergic neuronal differentiation and PSA-NCAM enrichment provides a useful system for the future study of hESCs.

subject areas

  • Animals
  • Biomarkers
  • Cell Differentiation
  • Cell Line, Tumor
  • Cell Shape
  • Coculture Techniques
  • Culture Media, Conditioned
  • Dopamine
  • Electrophysiology
  • Flow Cytometry
  • Gene Expression Profiling
  • Humans
  • Microarray Analysis
  • Molecular Sequence Data
  • Neural Cell Adhesion Molecules
  • Neurons
  • Stem Cells
  • Tyrosine 3-Monooxygenase
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Identity

International Standard Serial Number (ISSN)

  • 1547-3287

Digital Object Identifier (DOI)

  • 10.1089/scd.2005.14.517

PubMed ID

  • 16305337
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Additional Document Info

start page

  • 517

end page

  • 534

volume

  • 14

issue

  • 5

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