An optimized experimental strategy for efficient conversion of embryonic stem (ES)-derived mouse neural stem (NS) cells into a nearly homogeneous mature neuronal population

Neurobiol Dis. 2009 May;34(2):320-31. doi: 10.1016/j.nbd.2009.02.007. Epub 2009 Feb 21.

Abstract

NS cells are a homogeneous population of neural stem cells which were previously derived from embryonic stem cells as well as from the fetal and adult brain. Our previous reports have described a 21 day long neuronal differentiation protocol able to reproducibly convert adult SVZ-derived NS (aNS) cells into a population composed of 65% mature neurons and 35% glial cells. Here we have developed a different procedure specifically applicable to ES-derived NS cells in order to fully explore their neurogenic capacity. Differently from the aNS differentiation procedure, optimized neuronal output from ES-derived NS cells requires replating of the cells on appropriate substrates followed by sequential exposure to modified media. In these conditions, ES-derived NS cells differentiate into neurons with a barely appreciable quota of astrocytes and occasional oligodendrocytes. In particular, 21 days after the beginning of the treatment, 85% of the cells has differentiated into molecularly and electrophysiologically mature neurons belonging to the GABAergic lineage. The procedure, which is applicable with no considerable differences to different ES-derived NS cell lines and to NS cells at different passages, opens to the possibility of molecular and biochemical studies on close-to-uniform stem cell derived neurons.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Action Potentials / drug effects
  • Action Potentials / physiology
  • Animals
  • Astrocytes / cytology
  • Astrocytes / drug effects
  • Astrocytes / physiology
  • Biomarkers / analysis
  • Biomarkers / metabolism
  • Cell Culture Techniques
  • Cell Differentiation / drug effects
  • Cell Differentiation / physiology*
  • Cell Line
  • Cell Lineage / drug effects
  • Cell Lineage / physiology*
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / drug effects
  • Embryonic Stem Cells / physiology*
  • Interneurons / cytology
  • Interneurons / drug effects
  • Interneurons / physiology
  • Mice
  • Nerve Growth Factors / pharmacology
  • Nerve Tissue Proteins / analysis
  • Nerve Tissue Proteins / metabolism
  • Neurogenesis / drug effects
  • Neurogenesis / physiology*
  • Neurons / cytology
  • Neurons / physiology*
  • Oligodendroglia / cytology
  • Oligodendroglia / drug effects
  • Oligodendroglia / physiology
  • Sodium Channels / drug effects
  • Sodium Channels / physiology
  • Stem Cell Transplantation / methods
  • gamma-Aminobutyric Acid / metabolism

Substances

  • Biomarkers
  • Nerve Growth Factors
  • Nerve Tissue Proteins
  • Sodium Channels
  • gamma-Aminobutyric Acid