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Primary Cultures of Astrocytes and Neurons as Model Systems to Study the Metabolism and Metabolite Export from Brain Cells

  • Ketki Tulpule
  • Michaela C. Hohnholt
  • Johannes Hirrlinger
  • Ralf DringenEmail author
Protocol
Part of the Neuromethods book series (NM, volume 90)

Abstract

Primary cultures of astrocytes and neurons are frequently used to investigate metabolic properties of these two important brain cell types. Here we describe methods to generate primary cultures that are highly enriched in astrocytes or cerebellar granule neurons. These cultures are good model systems to investigate the basal metabolism of astrocytes and neurons as well as metabolite export from these cells. As examples for such studies, we describe here in detail the robust practical procedures that we use to investigate cellular glucose consumption and lactate production as well as the cellular contents and the export of glutathione from cultured brain cells. In this context we also describe some viability assays that are useful to confirm that a given experimental treatment is not toxic for cultured neural cells.

Key words

Astrocytes Metabolite export Glucose Glycolytic flux GSH Neurons Lactate Viability 

Notes

Acknowledgements

We very much like to thank Dr. Bernd Hamprecht (Tuebingen, Germany) and Dr. Mike Cousin (Edinburgh, Scotland) for the opportunities to learn the methods to prepare APCs and CGNCs, respectively, in their laboratories.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Ketki Tulpule
    • 1
    • 2
  • Michaela C. Hohnholt
    • 1
    • 2
  • Johannes Hirrlinger
    • 3
  • Ralf Dringen
    • 1
    • 2
    Email author
  1. 1.Centre for Biomolecular Interactions BremenUniversity of BremenBremenGermany
  2. 2.Centre for Environmental Research and Sustainable TechnologyBremenGermany
  3. 3.Carl-Ludwig-Institute for PhysiologyUniversity of LeipzigLeipzigGermany

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