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Assessing Cellular Hypoxic Status In Situ Within the Bone Marrow Microenvironment

  • Ute Suessbier
  • César Nombela-ArrietaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2017)

Abstract

Hematopoietic stem cells are maintained and regulated in spatially confined microenvironments within the bone marrow, in which oxygen availability is hypothesized to be very limited. The hypoxic nature of HSC niches is proposed to play a fundamental role in the preservation of fundamental stem cell properties through the induction of a distinct glycolytic metabolic profile in HSCs. Thus, the capacity to determine oxygen levels or cellular oxygenation status in specific tissue locations is essential to deepen our understanding of HSC biology. We here describe a methodology to indirectly quantify the hypoxic status of individual cells in situ within histological sections of bone marrow tissues. We employ the well-characterized nitroimidazole probe, pimonidazole, which acts as an oxygen mimetic and irreversibly incorporates into cellular proteins only under hypoxic conditions. The use of fluorescently labeled antibodies that recognize pimonidazole epitopes then enables the indirect assessment of the intracellular hypoxic status and its relationship to cell positioning within the complex tissue topography of the bone marrow.

Key words

Hypoxic status HSC niche Imaging Microenvironment Metabolic state 

Notes

Acknowledgments

This work was supported by the Swiss National Research Foundation (grant number 31003A_159597/1) and an FP7 Marie Curie Career Integration Grant (PCIG13-GA-2013- 618633) from the European Union.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Medical Oncology and HematologyUniversity of ZurichZurichSwitzerland

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