Detection of calcium transients in embryonic stem cells and their differentiated progeny

Jason S. Meyer, Gregory Tullis, Christopher Pierret, Kathleen M. Spears, Jason A. Morrison, Mark D. Kirk

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

A central issue in stem cell biology is the determination of function and activity of differentiated stem cells, features that define the true phenotype of mature cell types. Commonly, physiological mechanisms are used to determine the functionality of mature cell types, including those of the nervous system. Calcium imaging provides an indirect method of determining the physiological activities of a mature cell. Camgaroos are variants of yellow fluorescent protein that act as intracellular calcium sensors in transfected cells. We expressed one version of the camgaroos, Camgaroo-2, in mouse embryonic stem (ES) cells under the control of the CAG promoter system. Under the control of this promoter, Camgaroo-2 fluorescence was ubiquitously expressed in all cell types derived from the ES cells that were tested. In response to pharmacological stimulation, the fluorescence levels in transfected cells correlated with cellular depolarization and hyperpolarization. These changes were observed in both undifferentiated ES cells as well as ES cells that had been neurally induced, including putative neurons that were differentiated from transfected ES cells. The results presented here indicate that Camgaroo-2 may be used like traditional fluorescent proteins to track cells as well as to study the functionality of stem cells and their progeny.

Original languageEnglish (US)
Pages (from-to)1191-1203
Number of pages13
JournalCellular and Molecular Neurobiology
Volume29
Issue number8
DOIs
StatePublished - Dec 1 2009

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Keywords

  • Calcium imaging
  • Camgaroo
  • Fluorescent indicators
  • Stem cells

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Cell Biology

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