Abstract
Expansion of human stem cells before cell therapy is typically performed at 20% O2. Growth in these pro-oxidative conditions can lead to oxidative stress and genetic instability. Here, we demonstrate that culture of human mesenchymal stem cells at lower, physiological O2 concentrations significantly increases lifespan, limiting oxidative stress, DNA damage, telomere shortening and chromosomal aberrations. Our gene expression and bioenergetic data strongly suggest that growth at reduced oxygen tensions favors a natural metabolic state of increased glycolysis and reduced oxidative phosphorylation. We propose that this balance is disturbed at 20% O2, resulting in abnormally increased levels of oxidative stress. These observations indicate that bioenergetic pathways are intertwined with the control of lifespan and decisively influence the genetic stability of human primary stem cells. We conclude that stem cells for human therapy should be grown under low oxygen conditions to increase biosafety.
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Abbreviations
- ECAR:
-
extra cellular acidification rate
- FDR:
-
false discovery rate
- OCR:
-
oxygen consumption rate
- OXPHOS:
-
oxidative phosphorylation
- TRAP:
-
telomere repeat amplification protocol
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Acknowledgements
We are indebted to Dr. Judith Campisi (Buck Institute for Age Research) for providing the telomere length control cells and the hTert lentiviral vector, to Jose Manuel Ligos and the Cytometry Unit (CNIC) for advice on FACS analysis, Antonio Diez-Juan and Kenneth McCreath for critical reading of the paper, Marta Ramón (CNIC) for secretarial assistance and Simon Bartlett (CNIC) for editorial support. This work was supported by grants to AB from the Ministry of Science and Innovation (SAF 2008-02099; PLE2009-0147 and PSE-010000-2009-3), the Comunidad Autónoma de Madrid (P-BIO-0306-2006) and the Red de Terapia Celular del Instituto de Salud Carlos III (TerCel); to ES from the Fundación Mutua Madrileña, the Ministry of Education (Ramon y Cajal program), and the Ministry of Health (FIS PI071023) to JAE from the Ministry of Science and Innovation (SAF 2009-08007 and CSD2007-00020). JCE is currently a predoctoral fellow funded by TerCel. The CNIC is supported by the Spanish Ministry of Science and Innovation and the Pro-CNIC Foundation.
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Estrada, J., Albo, C., Benguría, A. et al. Culture of human mesenchymal stem cells at low oxygen tension improves growth and genetic stability by activating glycolysis. Cell Death Differ 19, 743–755 (2012). https://doi.org/10.1038/cdd.2011.172
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DOI: https://doi.org/10.1038/cdd.2011.172
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