A Comparative Study of Suspension Cultivation Systems for the Expansion of Undifferentiated Mouse Embryonic Stem Cells

  • Magda Tomala
  • Sabrina Schmeckebier
  • Ruth Olmer
  • Pierre Moretti
  • Frank Stahl
  • Ulrich Martin
  • Thomas Scheper
  • Cornelia Kasper
Conference paper
First Online:
Part of the ESACT Proceedings book series (ESACT, volume 5)

Abstract

Embryonic Stem (ES) cells are self- renewing and pluripotent cells that can differentiate into a variety of cell lineages. Implementation in tissue engineering or as model systems for drug discovery makes ES cells attractive as a cell source. To use ES cells for these applications, technologies are required to generate a large number of cells with defined characteristics. Currently, adherent culture methods are routinely applied for the maintenance of undifferentiated ES cells. However, perfused and stirred bioreactors enable a more homogenous environment and, more importantly, facilitate the ability to monitor and control culture parameters (e.g. oxygen content and pH) which is advantageous compared with static culture vessels. In this work, four different suspension cultivation systems (two spinner flasks with different stirrer design, an Erlenmeyer flask and a Petri dish) were compared with respect to the ability to generate undifferentiated ES cells. Therefore, the murine ES cell line E14.1, 129/Ola with an eGFP transgene targeted to the Brachyury locus was used. Besides the evaluation of biomass production, pluripotency marker expression was analyzed applying flow cytometry.

Keywords

Embryonic Stem Cell Spinner Flask Undifferentiated Embryonic Stem Cell Suspension Cultivation System Human Leukemia Inhibitory Factor 
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Notes

Acknowledgements

The authors would like to thank Gordon Keller, Toronto, for kindly providing transgenic Brachyury-eGFP ES cells. This work was supported by funding from the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) for the Cluster of Excellence REBIRTH.

Reference

  1. Majore I, Moretti P, Hass R, Kasper C. Identification of subpopulations in mesenchymal stem cell-like cultures from human umbilical cord. Cell Commun Signal. 2009, 7, 6.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Magda Tomala
    • 1
  • Sabrina Schmeckebier
    • 2
  • Ruth Olmer
    • 1
  • Pierre Moretti
    • 1
  • Frank Stahl
    • 1
  • Ulrich Martin
    • 2
  • Thomas Scheper
    • 1
  • Cornelia Kasper
    • 1
  1. 1.Institute of Technical Chemistry, Leibniz University of HannoverHannoverGermany
  2. 2.Department of Cardiac-, Thoracic-, Transplantation and Vascular SurgeryREBIRTH-Center for Regenerative Medicine, Hannover Medical SchoolHannoverGermany

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