Metabolism
Metabolism
Technical
Expert
The Metabolism Core Facility is an advanced research platform specialized in the investigation of cellular metabolism and bioenergetics.
The facility is composed of four specialized Infrastructure Units, that provide cutting-edge technologies and expert support for mitochondrial function analysis, in vivo and in vitro metabolic phenotyping, and both targeted and untargeted metabolomics.
Dr. Simone Reano
INSTRUMENTS
The facility is equipped with:
- Oroboros O2k High-Resolution Respirometry (Oroboros Instruments)
- Metabolic cages (OxyletPro)
- Seahorse XF Pro
- Thermo Scientific Q Exactive Plus Mass Spectrometer + Vanquish UHPLC Systems
- LECO Pegasus BT 4D GCxGC-TOF (SPME and HS)
- LECO HRT 4D GCxGC-TOF
To reserve instruments, use the “Agendo” management system: https://upo.agendoscience.com/.
Infrastucture units
The Metabolism Core Facility is composed of four Infrastructure Units:
1. High-Resolution Respirometry
The Oroboros O2k (O2k) is a unique high-resolution approach to monitor cellular and mitochondrial respiratory function. High-resolution respirometry (HRR) is befitting for the analysis of biopsies with limited amounts of sample (diagnosis of genetic and acquired mitochondrial defects in pediatric patients, diabetes), cell cultures with limited cell numbers or low activities, mutants with diminished respiratory capacity. It can also be used to measure chemical oxidation rates, antioxidant capacities, oxygen kinetic, and intracellular oxygen at physiological levels.
The versatility of O2k allows simultaneous measurement of respiration and fluorometric signals (H2O2 flux, mt-membrane potential, Ca2+, ADP-ATP phosphorylation). O2k is a relevant tool for the analysis of pathological effects causing reduced respiration, e.g., mitochondrial and metabolic diseases, ischemia-reperfusion injury, oxidative stress, apoptosis, and aging.
SCIENTIFIC SUPERVISOR
Prof. Nicoletta Filigheddu
The Unit offers:
- Support in experimental design.
- Identification of the most appropriate protocol for your experimental issue:
- Oxygen consumption rate (OCR) in intact cell.
- OCR in permeabilized cells and analysis of mitochondrial complex activity.
- OCR in permeabilized muscle fibers and in tissue homogenate.
- OCR in isolated mitochondria from tissue and cells.
- Mitochondrial H2O2 and ATP production in parallel to OCR.
- Mitochondrial Ca2+ flux in parallel to OCR.
- Experiment execution, data analysis, and graphical results (Excel or Prism).
2. Mouse metabolic phenotyping
The Unit supports and facilitates in vivo metabolic research by offering state-of-the-art equipment, as metabolic cages (OxyletPro), expert assistance for planning and performing experiments, with data analysis for all in vivo research. OxyletPro, an optimized system for studies in laboratory research models, utilizes indirect calorimetry to evaluate respiratory metabolism (VO2 consumption/VCO2 production).
SCIENTIFIC SUPERVISOR
Prof. Nicoletta Filigheddu
The Unit offers:
- Guidance for project planning and experiment design BEFORE THE PROJECT SUBMISSION TO OPBA/MINISTERO DELLA SALUTE for clearance.
- Support and access to specialized equipment including OxyletPro metabolic cages.
- Technical assistance to perform metabolic tests (glucose and insulin tolerance tests).
- Technical assistance for blood sampling, tissue extraction, and specialized surgical procedures.
- Support for data analysis and interpretation for Resting Energy Expenditure, Glucose Tolerance Test (oral or IP), or Insulin Tolerance Test assays.
3. Metabolic Phenotyping
This unit’s primary focus is on characterizing cellular metabolic phenotypes through the precise measurement of key metabolic parameters. Utilizing the Seahorse XF Pro analyzer, the unit enables detailed investigations into cellular respiration, glycolytic activity, and energy production pathways.
This capability is crucial for understanding cellular responses to various physiological and pathological conditions. The unit also allows for specialized assays, including the evaluation of mitochondrial complexes activities following cell permeabilization and the assessment of T cell activation status and bioenergetics. Notably, metabolic profiling is increasingly recognized as a critical tool in immuno-oncology, as metabolic reprogramming plays a key role in T cell activation, exhaustion, and the efficacy of CAR-T cell therapies and immune checkpoint inhibitors.
SCIENTIFIC SUPERVISOR
Prof. Alessio Menga
The Unit offers:
- Real-time assessment of oxygen consumption rate (OCR) to evaluate mitochondrial respiration (e.g., using the Seahorse XF Mito Stress Test).
- Measurement of extracellular acidification rate (ECAR) to quantify glycolytic flux (e.g., using the Seahorse XF Glycolysis Stress Test).
- Analysis of ATP production rates and metabolic flexibility (e.g., using the Seahorse XF ATP Rate Assay).
- Evaluation of mitochondrial complexes activities following cell permeabilization (Plasma Membrane Permeabilizer).
- Detailed assessment of T cell activation status and bioenergetics, with a specialized focus on: 1) Characterizing metabolic shifts associated with T cell activation, differentiation, and effector function; 2) Investigating metabolic vulnerabilities linked to T cell exhaustion and dysfunction in cancer and chronic infections; 3) Evaluating metabolic modulation strategies to enhance T cell persistence and antitumor activity in CAR-T cell therapies and to overcome resistance to immune checkpoint inhibitors.
- Design and execution of metabolic assays to study the impact of drugs, toxins, and genetic manipulations on cellular metabolism.
- Expert consultation on experimental design, data acquisition, and data interpretation.
4. Untargeted and targeted metabolomics
The unit offers comprehensive analyses of metabolites from diverse samples using advanced high- and medium-resolution mass spectrometers coupled to liquid chromatography, mono- (GC-MS) and bi-dimensional gas chromatography (GCxGC-MS).
The unit supports research on untargeted metabolomic profiling of biofluids (plasma, serum, CSF, urine, saliva, stools, etc.), tissues and cells using GCxGC-MS, allowing the relative quantification of hundreds of small molecules. For the quantification of thousands of lipid species, the analysis is performed using liquid chromatography. Some targeted methods (short chain fatty acids, neurotransmitters, amino acids, etc.) are also available.
If needed, the staff can develop new customized analytical methods and can perform data analysis, pathway analysis and omics data integration. The unit guides investigators through the experimental design and statistical considerations of their studies.
SCIENTIFIC SUPERVISOR
Prof. Marcello Manfredi
The Unit offers:
- Global untargeted metabolomic and lipidomic profiling with comparative analysis.
- Targeted metabolomic profiling.
- Profiling of biological samples.
- High throughput data analysis (including basic statistics and pathway analysis).
CONTACTS
For any information, please contact
corefacilities.ipazia@uniupo.it
For any information, please contact
corefacilities.ipazia@uniupo.it
or segreteria.caad@uniupo.
CAAD
Center for Translational Research on
Autoimmune and Allergic Disease
Centro di Ricerca Traslazionale
sulle Malattie Autoimmuni e Allergiche
Corso Trieste 15/A
Novara – 28100 (ITALY)
Metabolism
Metabolism
The Metabolism Core Facility is an advanced research platform specialized in the investigation of cellular metabolism and bioenergetics.
The facility is composed of four specialized Infrastructure Units, that provide cutting-edge technologies and expert support for mitochondrial function analysis, in vivo and in vitro metabolic phenotyping, and both targeted and untargeted metabolomics.
INSTRUMENTS
The facility is equipped with:
- Oroboros O2k High-Resolution Respirometry (Oroboros Instruments)
- Metabolic cages (OxyletPro)
- Seahorse XF Pro
- Thermo Scientific Q Exactive Plus Mass Spectrometer + Vanquish UHPLC Systems
- LECO Pegasus BT 4D GCxGC-TOF (SPME and HS)
- LECO HRT 4D GCxGC-TOF
To reserve instruments, use the “Agendo” management system: https://upo.agendoscience.com/.
Infrastucture units
The Metabolism Core Facility is composed of four Infrastructure Units:
1. High-Resolution Respirometry
The Oroboros O2k (O2k) is a unique high-resolution approach to monitor cellular and mitochondrial respiratory function. High-resolution respirometry (HRR) is befitting for the analysis of biopsies with limited amounts of sample (diagnosis of genetic and acquired mitochondrial defects in pediatric patients, diabetes), cell cultures with limited cell numbers or low activities, mutants with diminished respiratory capacity. It can also be used to measure chemical oxidation rates, antioxidant capacities, oxygen kinetic, and intracellular oxygen at physiological levels.
The versatility of O2k allows simultaneous measurement of respiration and fluorometric signals (H2O2 flux, mt-membrane potential, Ca2+, ADP-ATP phosphorylation). O2k is a relevant tool for the analysis of pathological effects causing reduced respiration, e.g., mitochondrial and metabolic diseases, ischemia-reperfusion injury, oxidative stress, apoptosis, and aging.
SCIENTIFIC SUPERVISOR
Prof. Nicoletta Filigheddu
The Unit offers:
- Support in experimental design.
- Identification of the most appropriate protocol for your experimental issue:
- Oxygen consumption rate (OCR) in intact cell.
- OCR in permeabilized cells and analysis of mitochondrial complex activity.
- OCR in permeabilized muscle fibers and in tissue homogenate.
- OCR in isolated mitochondria from tissue and cells.
- Mitochondrial H2O2 and ATP production in parallel to OCR.
- Mitochondrial Ca2+ flux in parallel to OCR.
- Experiment execution, data analysis, and graphical results (Excel or Prism).
2. Mouse metabolic phenotyping
The Unit supports and facilitates in vivo metabolic research by offering state-of-the-art equipment, as metabolic cages (OxyletPro), expert assistance for planning and performing experiments, with data analysis for all in vivo research. OxyletPro, an optimized system for studies in laboratory research models, utilizes indirect calorimetry to evaluate respiratory metabolism (VO2 consumption/VCO2 production).
SCIENTIFIC SUPERVISOR
Prof. Nicoletta Filigheddu
The Unit offers:
- Guidance for project planning and experiment design BEFORE THE PROJECT SUBMISSION TO OPBA/MINISTERO DELLA SALUTE for clearance.
- Support and access to specialized equipment including OxyletPro metabolic cages.
- Technical assistance to perform metabolic tests (glucose and insulin tolerance tests).
- Technical assistance for blood sampling, tissue extraction, and specialized surgical procedures.
- Support for data analysis and interpretation for Resting Energy Expenditure, Glucose Tolerance Test (oral or IP), or Insulin Tolerance Test assays.
3. Metabolic Phenotyping
This unit’s primary focus is on characterizing cellular metabolic phenotypes through the precise measurement of key metabolic parameters. Utilizing the Seahorse XF Pro analyzer, the unit enables detailed investigations into cellular respiration, glycolytic activity, and energy production pathways.
This capability is crucial for understanding cellular responses to various physiological and pathological conditions. The unit also allows for specialized assays, including the evaluation of mitochondrial complexes activities following cell permeabilization and the assessment of T cell activation status and bioenergetics. Notably, metabolic profiling is increasingly recognized as a critical tool in immuno-oncology, as metabolic reprogramming plays a key role in T cell activation, exhaustion, and the efficacy of CAR-T cell therapies and immune checkpoint inhibitors.
SCIENTIFIC SUPERVISOR
Prof. Alessio Menga
The Unit offers:
- Real-time assessment of oxygen consumption rate (OCR) to evaluate mitochondrial respiration (e.g., using the Seahorse XF Mito Stress Test).
- Measurement of extracellular acidification rate (ECAR) to quantify glycolytic flux (e.g., using the Seahorse XF Glycolysis Stress Test).
- Analysis of ATP production rates and metabolic flexibility (e.g., using the Seahorse XF ATP Rate Assay).
- Evaluation of mitochondrial complexes activities following cell permeabilization (Plasma Membrane Permeabilizer).
- Detailed assessment of T cell activation status and bioenergetics, with a specialized focus on: 1) Characterizing metabolic shifts associated with T cell activation, differentiation, and effector function; 2) Investigating metabolic vulnerabilities linked to T cell exhaustion and dysfunction in cancer and chronic infections; 3) Evaluating metabolic modulation strategies to enhance T cell persistence and antitumor activity in CAR-T cell therapies and to overcome resistance to immune checkpoint inhibitors.
- Design and execution of metabolic assays to study the impact of drugs, toxins, and genetic manipulations on cellular metabolism.
- Expert consultation on experimental design, data acquisition, and data interpretation.
4. Untargeted and targeted metabolomics
The unit offers comprehensive analyses of metabolites from diverse samples using advanced high- and medium-resolution mass spectrometers coupled to liquid chromatography, mono- (GC-MS) and bi-dimensional gas chromatography (GCxGC-MS).
The unit supports research on untargeted metabolomic profiling of biofluids (plasma, serum, CSF, urine, saliva, stools, etc.), tissues and cells using GCxGC-MS, allowing the relative quantification of hundreds of small molecules. For the quantification of thousands of lipid species, the analysis is performed using liquid chromatography. Some targeted methods (short chain fatty acids, neurotransmitters, amino acids, etc.) are also available.
If needed, the staff can develop new customized analytical methods and can perform data analysis, pathway analysis and omics data integration. The unit guides investigators through the experimental design and statistical considerations of their studies.
SCIENTIFIC SUPERVISOR
Prof. Marcello Manfredi
The Unit offers:
- Global untargeted metabolomic and lipidomic profiling with comparative analysis.
- Targeted metabolomic profiling.
- Profiling of biological samples.
- High throughput data analysis (including basic statistics and pathway analysis).
CONTACTS
For any information, please contact
corefacilities.ipazia@uniupo.it
For any information, please contact
corefacilities.ipazia@uniupo.it
or segreteria.caad@uniupo.
CAAD
Center for Translational Research on
Autoimmune and Allergic Disease
Centro di Ricerca Traslazionale
sulle Malattie Autoimmuni e Allergiche
Corso Trieste 15/A
Novara – 28100 (ITALY)