Differentiation of rhesus embryonic stem cells to neural progenitors and neurons

John D. Calhoun, Nevin A Lambert, Maya M. Mitalipova, Scott A. Noggle, Ian Lyons, Brian G. Condie, Steven L. Stice

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Embryonic stem (ES) cells are pluripotent cells capable of differentiating into cell lineages derived from all primary germ layers including neural cells. In this study we describe an efficient method for differentiating rhesus monkey ES cells to neural lineages and the subsequent isolation of an enriched population of Nestin and Musashi positive neural progenitor (NP) cells. Upon differentiation, these cells exhibit electrophysiological characteristics resembling cultured primary neurons. Embryoid bodies (EBs) were formed in ES growth medium supplemented with 50% MEDII. After 7 days in suspension culture, EBs were transferred to adherent culture and either differentiated in serum containing medium or expanded in serum free medium. Immunocytochemistry on differentiating cells derived from EBs revealed large networks of MAP-2 and NF200 positive neurons. DAPI staining showed that the center of the MEDII-treated EBs was filled with rosettes. NPs isolated from adherent EB cultures expanded in serum free medium were passaged and maintained in an undifferentiated state by culture in serum free N2 with 50% MEDII and bFGF. Differentiating neurons derived from NPs fired action potentials in response to depolarizing current injection and expressed functional ionotropic receptors for the neurotransmitters glutamate and γ-aminobutyric acid (GABA). NPs derived in this way could serve as models for cellular replacement therapy in primate models of neurodegenerative disease, a source of neural cells for toxicity and drug testing, and as a model of the developing primate nervous system.

Original languageEnglish (US)
Pages (from-to)191-197
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume306
Issue number1
DOIs
StatePublished - Jun 20 2003

Fingerprint

Embryoid Bodies
Embryonic Stem Cells
Stem cells
Neurons
Serum-Free Culture Media
Neurodegenerative diseases
Aminobutyrates
Nestin
Neurotransmitter Receptor
Primates
Neurology
Toxicity
Glutamic Acid
Suspensions
Ionotropic Glutamate Receptors
Germ Layers
Cell Lineage
Drug-Related Side Effects and Adverse Reactions
Macaca mulatta
Serum

Keywords

  • Electrophysiology
  • Embryonic stem cell
  • Human
  • Neural progenitor
  • Neural stem cell
  • Neuron
  • Primate

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Differentiation of rhesus embryonic stem cells to neural progenitors and neurons. / Calhoun, John D.; Lambert, Nevin A; Mitalipova, Maya M.; Noggle, Scott A.; Lyons, Ian; Condie, Brian G.; Stice, Steven L.

In: Biochemical and Biophysical Research Communications, Vol. 306, No. 1, 20.06.2003, p. 191-197.

Research output: Contribution to journalArticle

Calhoun, John D. ; Lambert, Nevin A ; Mitalipova, Maya M. ; Noggle, Scott A. ; Lyons, Ian ; Condie, Brian G. ; Stice, Steven L. / Differentiation of rhesus embryonic stem cells to neural progenitors and neurons. In: Biochemical and Biophysical Research Communications. 2003 ; Vol. 306, No. 1. pp. 191-197.
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