Hypoxia Core

The Hypoxia Core of the IU School of Medicine Cooperative Center of Excellence in Hematology (IU-CCEH) provides resources for members of the IU-CCEH center, other centers, and outside investigators in the area of nonmalignant hematology to generate information critical for better understanding cellular, molecular, and biochemical aspects of hematopoietic stem, progenitor, immune and stromal cells under relevant physiological conditions (i.e., lowered oxygen tension). 

O2 tension within bone marrow (BM;1-5%) and cord blood (CB) or mobilized peripheral blood (mPB; <10%), is lower than in the ambient air (~21% O2) in which they are routinely collected for analysis. While it has been known for >40 years that HSC and HPC grow better ex-vivo in lowered (≤ 5%) O2, the core's report (Mantel, et. al., Cell, 2015) and more recent work demonstrate that collection/processing of BM, CB, and mPB in ambient air results in large decreases in phenotypically defined and functional HSC and increased HPC numbers within minutes of cell exposure to ambient air. The core termed this process Extra Physiological Shock/Stress (EPHOSS). Collection/processing of cells at 3% O2, such that they are never exposed to ambient air, resulted in two to five-fold increases in phenotypically- and functionally-detectable HSC. As well, this manifested as different gene expression patterns and responsiveness to stimuli associated with HSC. Similar changes were also noted with murine pre-leukemic and immune cells. Reevaluation of hematopoietic function associated with maintenance of HSC and HPC at lowered O2 levels can now be elucidated through the expertise of the Hypoxia Core. New data derived with the core's hypoxia chamber demonstrates the incredible power of the core to serve the hematopoietic community.

Specific aims of the Hypoxia Core involving normal and nonmalignant murine and human hematology are:

  • Provide outstanding, consistent and timely analysis of mouse and human BM, mPB, immune and stromal cells, and human CB, BM, and mPB collected/processed at 3%, as well as other, O2 tensions compared to that in ambient air.
  • Coordinate with the other IU-CCEH cores for in-depth analysis of cells collected in hypoxia vs. ambient air.
  • Enhance productivity of IU-CCEH and other CCEH members, and outside investigators by providing a centralized set of services with reduced cost, and expert guidance not currently available elsewhere.
  • Provide advice regarding intricacies of such studies, analyses, and overcoming potential problems. The core provides an economic resource that will add new and significant cutting-edge scientific analysis to greatly advance our current understanding of HSC/HPC and immune and stromal cell biology in ways that will increase their translational use for clinical benefit.

Services

  • Expert guidance on designing experimental protocols.
  • Options to have the core staff train individuals to use the equipment or have our trained staff perform the experiment.
  • Access to an Agilent Seahorse machine under varying O2 and CO2 concentrations to examine cell metabolism under physiological conditions.

Equipment

diagram of the modified biospherix working station

Modified BioSpherix Working Station

Each chamber and buffer station can be set at different chamber pressures, O2 and CO2 concentrations independent of one another. Sensors in each chamber monitor O2 and CO2 concentrations, humidity, chamber pressure and temperature. 

coy lab products station

Coy Lab Products Station

The Coy Lab Products Station allows for injection of cells into mice under modified O2 and CO2 conditions. Mice have access to room air pumped into a nose cone while their tail or skin is exposed to lowered O2 or CO2 conditions.

Hypoxia Core Policy and Procedures for Acknowledgment, Authorship, and Collaboration

All research work performed by the Hypoxia Core should be acknowledged in all ensuing publications. Fees paid for services provided by the core should not negate the potential of co-authorship by core scientists. These acknowledgements and achievements are important for the existence and continued funding of the core facility. Given that many validation procedures require advanced intellectual involvement by the core’s scientists, it is expected that this acknowledgement will be in the form of co-authorship according to the guidelines for authorship recommended by the International Committee of Medical Journal Editors.

Hypoxia core scientists performing one or more of the following tasks meet the criteria for authorship:

  • Literature review and providing assistance in selecting/conceiving the experiments appropriate for the study goals
  • Experimental design
  • Assay development/optimization
  • Data analysis and interpretation of data, or both
  • Preparation of pertinent sections/figures for the manuscript and revisions to address reviewer concerns/questions

All contributions that do not meet the criteria for authorship should be listed in the acknowledgments section. Examples of the tasks requiring an appropriate mention in the acknowledgment section are:

  • Mere collection of data using PI-provided protocols that require no further optimization and/or trouble-shooting
  • Mere collection of data that requires no data analysis or interpretation by core scientists

Contact the Hypoxia Core

For questions or inquiries about usage of the core, please contact Maegan L. Capitano, PhD.

Email Dr. Capitano

Publications using Hypoxia Core equipment and services

Kumar B, Adebayo AK, Prasad M, Capitano ML, Wang R, Bhat-Nakshatri P, Anjanappa M, Simpson E, Chen D, Liu Y, Schilder JM, Colter AB, Maguire C, Temm CJ, Sandusky G, Doud EH, Wijeratne AB, Mosley AL, Broxmeyer HE, Nakshatri H.  Tumor tissue collection and processing under physoxia allows highly relevant detection of signaling networks and drug sensitivity of cancer cells. Science Advances.  2021. In press.

Noonan ML, Ni P, Agoro R, Sacks SA, Swallow EA, Wheeler JA, Clinkenbeard EL, Capitano ML, Prideaux M, Atkins GJ, Thompson WR, Allen MR, Broxmeyer HE, White KE. The HIF-PHI BAY 85-3934 (Molidustat) Improves Anemia and Is Associated With Reduced Levels of Circulating FGF23 in a CKD Mouse Model. J Bone Miner Res. 2021 Jun;36(6):1117-1130. doi: 10.1002/jbmr.4272. Epub 2021 Mar 10. PMID: 33592127; PMCID: PMC8255270.

Capitano ML, Mohamad SF, Cooper S, Guo B, Huang X, Gunawan AM, Sampson C, Ropa J, Srour EF, Orschell CM, Broxmeyer HE. Mitigating oxygen stress enhances aged mouse hematopoietic stem cell numbers and function. J Clin Invest. 2021 Jan 4;131(1):e140177. doi: 10.1172/JCI140177. PMID: 33393491; PMCID: PMC7773345.

Aljoufi A, Cooper S, Broxmeyer HE. Collection and Processing of Mobilized Mouse Peripheral Blood at Lowered Oxygen Tension Yields Enhanced Numbers of Hematopoietic Stem Cells. Stem Cell Rev Rep. 2020 Oct;16(5):946-953. doi: 10.1007/s12015-020-10021-w. PMID: 32748332; PMCID: PMC7484397.

Broxmeyer HE, Capitano ML, Cooper S, Potchanant ES, Clapp DW. Numbers of long-term hematopoietic stem cells from bone marrow of fanca and fancc knockout mice can be greatly enhanced by their collection and processing in physioxia conditions. Blood Cells Mol Dis. 2021 Feb;86:102492. doi: 10.1016/j.bcmd.2020.102492. Epub 2020 Aug 28. PMID: 32896825; PMCID: PMC7686233.

Shin DY, Huang X, Gil CH, Aljoufi A, Ropa J, Broxmeyer HE. Physioxia enhances T-cell development ex vivo from human hematopoietic stem and progenitor cells. Stem Cells. 2020 Nov;38(11):1454-1466. doi: 10.1002/stem.3259. Epub 2020 Aug 15. PMID: 32761664; PMCID: PMC7901063.

Mantel CR, O'Leary HA, Chitteti BR, Huang X, Cooper S, Hangoc G, Brustovetsky N, Srour EF, Lee MR, Messina-Graham S, Haas DM, Falah N, Kapur R, Pelus LM, Bardeesy N, Fitamant J, Ivan M, Kim KS, Broxmeyer HE. Enhancing Hematopoietic Stem Cell Transplantation Efficacy by Mitigating Oxygen Shock. Cell. 2015 Jun 18;161(7):1553-65. doi: 10.1016/j.cell.2015.04.054. Epub 2015 Jun 11. PMID: 26073944; PMCID: PMC4480616.