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The dark side of BrdU in neural stem cell biology: detrimental effects on cell cycle, differentiation and survival

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Abstract

5-Bromo-2′-deoxyuridin (BrdU) is frequently used in anaylsis of neural stem cell biology, in particular to label and to fate-map dividing cells. However, up to now, only a few studies have addressed the question as to whether BrdU labeling per se affects the cells to be investigated. Here, we focused on the potential impact of BrdU on neurosphere cultures derived from the adult rat brain and on proliferation of progenitors in vivo. In vitro, neurospheres were pulsed for 48 h with BrdU, and cell proliferation, cell cycle, differentiation, survival and adhesion properties were subsequently analyzed. BrdU inhibited the expansion of neural progenitors as assessed by MTS assay and increased the fraction of cells in the G0/G1-phase of the cell cycle. Moreover, BrdU increased cell death and dose-dependently induced adherence of NPCs. Cell adherence was accompanied by a reduced amount of active matrix-metalloproteinase-2 (MMP-2). Furthermore, BrdU repressed neuronal and oligodendroglial differentiation, whereas astroglial fate was not affected. In contrast to the in vitro situation, BrdU apparently did not influence endogenous proliferation of NPCs or neurogenesis in concentrations that are typically used for labeling of neural progenitors in vivo. Our results reveal so far uncharacterized effects of BrdU on adult NPCs. We conclude that, because of its ubiquitous use in stem cell biology, any potential effect of BrdU of NPCs has to be scrutinized prior to interpretation of data.

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Authors and Affiliations

  1. Institute of Molecular Regenerative Medicine, Paracelsus Medical University Salzburg, Strubergasse 21, 5020, Salzburg, Austria

    Bernadette Lehner, Julia Marschallinger, Tanja Furtner, Sebastien Couillard-Despres, Francisco J. Rivera & Ludwig Aigner

  2. Department of Neurology, University of Regensburg, Regensburg, Germany

    Bernadette Lehner, Beatrice Sandner & Norbert Weidner

  3. Department of Organismic Biology, Division of Zoology and Functional Anatomy, University of Salzburg, Salzburg, Austria

    Christine Lehner & Hans-Christian Bauer

  4. Applied Cell Biology, Paracelsus Medical University Salzburg, Salzburg, Austria

    Christine Lehner & Hans-Christian Bauer

  5. Clinic of Gynaecology and Obstetrics, Caritas Hospital St. Josef, University of Regensburg, Regensburg, Germany

    Gero Brockhoff

  6. Clinic of Paraplegiology, University of Heidelberg, Heidelberg, Germany

    Norbert Weidner

Authors
  1. Bernadette Lehner
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  2. Beatrice Sandner
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  4. Christine Lehner
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  6. Sebastien Couillard-Despres
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  7. Francisco J. Rivera
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  8. Gero Brockhoff
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  9. Hans-Christian Bauer
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  10. Norbert Weidner
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  11. Ludwig Aigner
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Correspondence to Ludwig Aigner.

Additional information

The authors would like to thank the following funding agencies for their support: the Bavarian State Ministry of Sciences, Research and the Arts (ForNeuroCell grant), the Germany Federal Ministry of Education and Research (BMBF grants #0312134, #01GG0706 and #01GN0505), the EU-FP6-Programm "DiMI", LSHB-CT-2005-512146, and the state government of Salzburg.

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Lehner, B., Sandner, B., Marschallinger, J. et al. The dark side of BrdU in neural stem cell biology: detrimental effects on cell cycle, differentiation and survival. Cell Tissue Res 345, 313–328 (2011). https://doi.org/10.1007/s00441-011-1213-7

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  • DOI: https://doi.org/10.1007/s00441-011-1213-7

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