Dynamic mass redistribution assay decodes differentiation of a neural progenitor stem cell

J Biomol Screen. 2012 Oct;17(9):1180-91. doi: 10.1177/1087057112455059. Epub 2012 Aug 10.

Abstract

Stem cells hold great potential in drug discovery and development. However, challenges remain to quantitatively measure the functions of stem cells and their differentiated products. Here, we applied fluorescent imaging, quantitative real-time PCR, and label-free dynamic mass redistribution (DMR) assays to characterize the differentiation process of the ReNcell VM human neural progenitor stem cell. Immunofluorescence imaging showed that after growth factor withdrawal, the neuroprogenitor stem cell was differentiated into dopaminergic neurons, astrocytes, and oligodendrocytes, thus creating a neuronal cell system. High-performance liquid chromatography analysis showed that the differentiated cell system released dopamine upon depolarization with KCl. In conjunction with quantitative real-time PCR, DMR assays using a G-protein-coupled receptor agonist library revealed that a subset of receptors, including dopamine D(1) and D(4) receptors, underwent marked alterations in both receptor expression and signaling pathway during the differentiation process. These findings suggest that DMR assays can decode the differentiation process of stem cells at the cell system level.

MeSH terms

  • Astrocytes / cytology
  • Astrocytes / metabolism
  • Cells, Cultured
  • Chromatography, High Pressure Liquid
  • Dopamine / analysis
  • Dopaminergic Neurons / cytology
  • Dopaminergic Neurons / metabolism
  • Fluorescent Antibody Technique / methods
  • Humans
  • Molecular Imaging / methods*
  • Neural Stem Cells / cytology
  • Neural Stem Cells / drug effects
  • Neural Stem Cells / metabolism*
  • Neurogenesis*
  • Oligodendroglia / cytology
  • Oligodendroglia / metabolism
  • Potassium Chloride / pharmacology
  • Real-Time Polymerase Chain Reaction / methods*
  • Receptors, Dopamine D1 / metabolism
  • Receptors, Dopamine D4 / metabolism
  • Receptors, G-Protein-Coupled / agonists
  • Receptors, G-Protein-Coupled / metabolism
  • Signal Transduction / physiology
  • Small Molecule Libraries / pharmacology

Substances

  • DRD4 protein, human
  • Receptors, Dopamine D1
  • Receptors, G-Protein-Coupled
  • Small Molecule Libraries
  • Receptors, Dopamine D4
  • Potassium Chloride
  • Dopamine