The Cellular Health and Metabolism Facility supports a range of research projects across multiple research disciplines at UoB, including Exercise Metabolism and Nutrition, Translational Muscle Physiology, Pharmaceutical Nanoscience, Biomedical Science, Inflammation and Aging, Immunology and Immunotherapy, Microbiology and Infection, Cardiovascular Sciences, and Cancer Sciences. The facility also supports many collaborative research projects with other HE institutions and the pharmaceutical industry.
Ongoing research projects
Optimising XF analysis to metabolically profile human peripheral blood mononuclear cells (PBMCs) isolated from fresh and day-old blood
Primary Investigators - Dr Alex Wadley and Dr Jonathan Barlow
This project aims to explore a new protocol for metabolically profiling human PBMCs isolated from fresh and day-old whole blood using different isolation methods. Our experimental design will enable parameters of both mitochondrial and glycolytic activity to be examined from the same cells by measuring oxygen consumption rates and proton efflux rates, respectively, in the same well of an XFe96 microplate using XF analysis. Detailed analysis of XF data will provide key information on mitochondrial function, glycolytic activity, ATP supply flux, bioenergetic scope, supply flexibility index and the mitochondrial health index.
Probing the effects of exercise on human blood derived immune cell bioenergetics
Primary Investigators -: Dr Alex Wadley and Dr Jonathan Barlow
This project aims to explore the effect of exercise on the metabolic function of quiescent and activated T-cells and/or PBMCs isolated from human blood. T-cells and/or PBMCS will be isolated from resting and immediately or 2 hour post exercise blood samples. Following isolation, immune cells of interest will be washed into XF assay buffer and immobilised in XFe96 microplates. The effect of exercise on cellular bioenergetics will be examined in quiescent (inactivated) and activated cells using XF analysis combined with our metabolic profiling test.
Using XF technology to probe human neutrophil function
Primary Investigators: Dr Jonathan Barlow and Dr Aaron Scott
This project aims to explore the use of XF analysis to probe cellular bioenergetics and oxidative burst activity by human blood derived neutrophils. As previously published, XF technology can be used to examine the kinetics of neutrophil oxidative burst in real-time when activated with PMA. However, it is not very well understood if XF analysis can be used to probe the kinetics of other (more physiologically relevant) oxidative burst activators. Moreover, it is not clear to what extent XF analysis can be used to accurately probe glycolytic activity and/or mitochondrial function of human neutrophils. Using standardised metabolic assays including the mito-stress test and glycolytic rate assay as well as our own modified assay designs, this project will confirm which aspects of neutrophil function can be accurately explored using XF analysis.
Investigating the Role of Macrophage Metabolism during Inflammation
Primary Investigators: Dr Samuel Lara Reyna and Dr Jonathan Barlow
Dr. Lara Reyna's research focuses on investigating the role of macrophage metabolism in the inflammatory response, with a particular emphasis on macrophage activation and consequent metabolic effects. In collaboration with Dr Reyna, we are using Seahorse XF analysis to measure macrophage bioenergetics to determine the effects of different stimuli on macrophage metabolism. The overall aim of this project is to better understand the intricate interplay between metabolic changes of macrophage activation during inflammation.
Our research impact
Publications
For our most up to date manuscript contributions see Publications.
Conferences
Oral presentations
MitOX 2019. Measuring mitochondrial function in cells and tissues using high resolution respirometry: Exploring the involvement of mitochondria in the effects of exercise on insulin secretion by pancreatic beta-cells. Dr Jonathan Barlow
Metabolic Research and Cell-Based Assays User Meeting: Cancer Research UK’s Cambridge Institute 2020. Discover the power of your spheroids using extracellular flux analysis. Dr Jonathan Barlow
Poster presentations
BTS Discovery Toxicology 2022: Science-led Strategies for Success in Discovery & Development Toxicology. An optimised protocol to explore mitochondrial energy metabolism of single spheroids using Agilent Seahorse Extracellular Flux Analysis. N. J. Coltman, G. Rochford, N. J. Hodges, H. Ali-Boucetta, J. P. Barlow
Webinars
Agilent Technologies (April 26, 2022). Exploring Mitochondrial Energy Metabolism of 3D Microtissue Spheroids using Agilent Seahorse Extracellular Flux Analysis. This webinar discusses details for probing mitochondrial energy metabolism in single 3D spheroids using spheroid microplates with the XFe96 XF analyzer. Using different cancer cell lines, XF technology is demonstrated to be capable of distinguishing between cellular respiration in 3D spheroids of not only different sizes but also different volumes, cell numbers, DNA content and type. Dr Jonathan Barlow
What our researchers say
“The Cellular Health and Metabolism Facility has assisted my PhD research project which involves studying a novel metabolic phenotype we have observed in knockout cells. The facility has been incredibly supportive; providing the facilities, equipment and most importantly the expertise and knowledge needed to investigate this phenotype."
- Miss Carys Howell (PhD student, Dr Cunningham’s Lab)
“I cannot speak highly enough of the Cellular Health and Metabolism Facility at the University of Birmingham. This scientific hub has exceeded all of my expectations. The staff are exceptionally knowledgeable, professional, and dedicated to their work, making the centre a hub for scientific innovation and discovery. The facilities have cutting-edge equipment and laboratory consumables that enable researchers to conduct their work at the highest level. I am particularly overwhelmed with the commitment of the centre towards collaboration, which creates a supportive and engaging environment for everyone. Thanks for your hard work and dedication!" - Dr Samuel Lara Reyna (Postdoctoral Research Fellow, Dr Eva Frickel's Lab)