Call 2016 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB) Main Applicant Francesca Cutruzzolà (Dept.Biochemical Sciences-DSB) Co-Applicants Andrea Bellelli (DSB), Rita Mancini (Dept. Clinical and Molecular Medicine, DMCM)
Cell behaviour depends on environmental oxygen Oncology HYPEROXIA PHYSIOXIA HYPOXIA Immunity and inflammation Re-shaping Neurobiology CELLULAR METABOLISM Regenerative medicine Host-Pathogen interaction and GENE EXPRESSION Drug discovery Toxicology Environmental biology 13/05/2019 Page 2 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
pO2 (20,10,5-3%) Biological sample Cellular Metabolism Gene expression LIVING CELLS Core Checkpoint Gene Set Oxygen Consumption rate (CCGS) Glycolitic activity hif, idh, shmt, myc … .. ATP production Specific Gene Set Carbon source (SGS) Custom-made (glucose, glutamine, fatty acids) 13/05/2019 Page 3 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
NUMBERS AND FACTS THE APPLICATION THE LAB • 35 m 2 dedicated renovated lab Supporting Staff Members: • Department of Biochemical Sciences 52 members (main campus) building CU027, rooms S32/S33 (ground floor). 8 Departments Involved: THE PERSONNEL Scienze Biochimiche Prof. Francesca Cutruzzolà Biologia e biotecnologie Darwin (Full Professor - Managing Director and Medicina clinica e molecolare Scientific Officer ) Medicina molecolare Prof. Serena Rinaldo Scienze e biotecnologie medico- (Associate Professor - Data analysis Manager ) chirurgiche Dr. Alessio Paone Medicina Sperimentale (RTD-A – Cell biologist ) Biologia Ambientale Dr. Giovanna Boumis (Technician – technical support ) Fisiologia e farmacologia 13/05/2019 Page 4 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
UNIQUE IN ITALY Seahorse analyser Dedicated Hypoxic chamber Cell Biology Molecular Biology 13/05/2019 Page 5 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
METABOLIC EXPERIMENT FLOWCHART Exp-1 Exp-2 Exp-3 Exp-4 SELECTION of ACCESS to the Equipment DATA storage methodology ASSISTANCE with operation EXPERIMENTAL design SUPPLY reagents and materials METHOD DEVELOPMENT 13/05/2019 Page 6 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
METABOLIC EXPERIMENT FLOWCHART Exp-1 Exp-2 Exp-3 Exp-4 SELECTION of ACCESS to the Equipment DATA storage methodology ASSISTANCE with operation EXPERIMENTAL design SUPPLY reagents and materials METHOD DEVELOPMENT 13/05/2019 Page 7 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
METABOLIC EXPERIMENT FLOWCHART Exp-1 Exp-2 Exp-3 Exp-4 SELECTION of ACCESS to the Equipment DATA storage methodology ASSISTANCE with operation EXPERIMENTAL design SUPPLY reagents and materials METHOD DEVELOPMENT 13/05/2019 Page 8 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
METABOLIC EXPERIMENT FLOWCHART Exp-1 Exp-2 Exp-3 Exp-4 SELECTION of ACCESS to the Equipment DATA storage methodology ASSISTANCE with operation EXPERIMENTAL design SUPPLY reagents and materials METHOD DEVELOPMENT 13/05/2019 Page 9 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
REPORTS TECHNICAL REPORT DETAILED SCIENTIFIC REPORT 13/05/2019 Page 10 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
Biological samples: examples from ongoing experiments DRUGS TRANSFECTIONS Normal eukaryotic cells Cancer cell lines (Fibroblasts, prostate cells DIFFERENTIATION (Leukemias, Lung adenocarcinoma, microglia ) Prostate and Cervical cancer..) STIMULATION WITH COMPOUNDS Bacteria Neurospheres (neural stem cells) 13/05/2019 Page 11 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
USER POLICIES AND GUIDELINES STEP 1 : Access is requested by e-mail to the Managing director or a delegate (see contacts). • Experimental background, number of samples to be assayed…. STEP 2 : Evaluation of the request by the HypACB team . Criteria: • Feasibility of the experiment and type of access required. Scheduled priority : Level 1: Internal academic users. Level 2: External academic users. Level 3: External non-academic investigators. • STEP 3 : Experiment is run by the platform personnel . The results of the experiments will be provided with all experimental details […]. Raw data will be stored as a copy for 5 years, according to international rules for scientific correctness. Data analysis can be performed by the end-users or by the HypACB team based on specific agreement. 13/05/2019 Page 12 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
USER POLICIES AND GUIDELINES STEP 1 : Access is requested by e-mail to the Managing director or a delegate (see contacts). • Experimental background, number of samples to be assayed…. STEP 2 : Evaluation of the request by the HypACB team . Criteria: • Feasibility of the experiment . Scheduled priority : Level 1: Internal academic users. Level 2: External academic users. Level 3: External non-academic investigators. • STEP 3 : Experiment is run. Type of access by the HypACB personnel . Acknowledgements Investigators are required to acknowledge the Hyp-ACB platform (and Sapienza Financial Contribution by Grant Number ) in any publication that results from the use of the platform. Names of individuals from platform will be included, if previously agreed or if they provided any intellectual input or additional effort. 13/05/2019 Page 13 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
USER POLICIES AND GUIDELINES STEP 1 : Access is requested by e-mail to the Managing director or a delegate (see contacts). • Experimental background, number of samples to be assayed….. STEP 2 : Evaluation of the request by the HypACB team . Criteria: • Feasibility of the experiment and type of access required. Scheduled priority : Level 1: Internal academic users. Level 2: External academic users. Level 3: External non-academic investigators. • STEP 3 : Type of access is agreed and Experiment is run. Acknowledgements Investigators are required to acknowledge the Hyp-ACB platform (and Sapienza Financial Contribution by Grant Number ) in any publication that results from the use of the platform. Names of individuals from platform will be included, if previously agreed or if they provided any intellectual input or additional effort. 13/05/2019 Page 14 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
USER POLICIES AND GUIDELINES STEP 1 : Access is requested by e-mail to the Managing director or a delegate (see contacts). • Experimental background, number of samples to be assayed…….. STEP 2 : Evaluation of the request by the HypACB team . Criteria: • Feasibility of the experiment . Scheduled priority : Level 1: Internal academic users. Level 2: External academic users. Level 3: External non-academic investigators. • STEP 3 : Type of access is agreed and Experiment is run. Results are provided and stored. Acknowledgements Investigators are required to acknowledge the Hyp-ACB platform (and Sapienza Financial Contribution by Grant Number ) in any publication that results from the use of the platform. Names of individuals from platform will be included, if previously agreed or if they provided any intellectual input or additional effort. 13/05/2019 Page 15 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
TYPE OF ACCESS Full Experiment Direct access experimental (from cell by end users planning and culture to data) follow-up 13/05/2019 Page 16 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
TYPE OF ACCESS Analysis Time Interpretation Materials Time allocation Method Personnel development 13/05/2019 Page 17 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
TYPE OF ACCESS: Service only Example of Experimental Costs (Metabolism 96-well plate) +35% for hypoxia 150 € 1200€ Analysis Time Interpretation Time allocation Materials Method Personnel development OFFICIAL PRICE LIST 2019 UNDER APPROVAL FOR SAPIENZA USERS: COST REDUCTION IS PLANNED (APPROX. 30-50% ) 13/05/2019 Page 18 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
TYPE OF ACCESS: Collaboration Example of Experimental Cost CHARGE REDUCTION (Metabolism 96-well plate) +35% for hypoxia 150€ 1200€ Analysis Time Interpretation Time allocation Materials Method Personnel development OFFICIAL PRICE LIST UNDER APPROVAL 13/05/2019 Page 19 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
TYPE OF ACCESS: Training Experiment Access only Full and access 13/05/2019 Page 20 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
NEW REQUESTS WELLCOME! TEMPORARY WEBSITE www.macinmec/hypacb-facility/ 13/05/2019 Page 24 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
CONTACTS • Managing director: prof. Francesca Cutruzzolà francesca.cutruzzola@uniroma1.it Tel. 0649910955 Platform Staff Data Manager: Prof. Serena Rinaldo serena.rinaldo@uniroma1.it Cell Biology: Dr. Alessio Paone alessio.paone@uniroma1.it Technical staff: Dr. Giovanna Boumis giovanna.boumis@uniroma1.it 13/05/2019 Page 25 Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
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