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Main Applicant Francesca Cutruzzolà (Dept.Biochemical Sciences-DSB)
Co-Applicants Andrea Bellelli (DSB), Rita Mancini (Dept. Clinical and Molecular Medicine, DMCM)
Platform for the Hypoxic Analysis of Cell Behaviour (Hyp-ACB)
Call 2016
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HYPEROXIA HYPOXIA
Cellbehaviourdependsonenvironmentaloxygen
Oncology Immunity and inflammation Neurobiology Regenerative medicine Host-Pathogen interaction Drug discovery Toxicology Environmental biology
Re-shapingCELLULARMETABOLISM
andGENEEXPRESSION
PHYSIOXIA
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Cellular Metabolism Gene expression
LIVING CELLS Oxygen Consumption rate
Glycolitic activity ATP production Carbon source
(glucose, glutamine, fatty acids)
Core Checkpoint Gene Set
(CCGS) hif, idh, shmt, myc…..
Specific Gene Set (SGS) Custom-made
Biologicalsample
pO2(20,10,5-3%)
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THEAPPLICATION
SupportingStaffMembers:52members
8DepartmentsInvolved:
ScienzeBiochimicheBiologiaebiotecnologieDarwinMedicinaclinicaemolecolare
MedicinamolecolareScienzeebiotecnologiemedico-
chirurgicheMedicinaSperimentaleBiologiaAmbientale
Fisiologiaefarmacologia
THELAB• 35m2dedicatedrenovatedlab
• DepartmentofBiochemicalSciences(maincampus)buildingCU027,roomsS32/S33(groundfloor).
THEPERSONNELProf.FrancescaCutruzzolà(FullProfessor-ManagingDirectorandScientificOfficer)Prof.SerenaRinaldo(AssociateProfessor-DataanalysisManager)Dr.AlessioPaone(RTD-A–Cellbiologist)Dr.GiovannaBoumis(Technician–technicalsupport)
NUMBERSANDFACTS
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CellBiology
SeahorseanalyserDedicatedHypoxicchamber
MolecularBiology
UNIQUE IN ITALY
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Exp-1 Exp-2 Exp-3 Exp-4
ACCESS to the Equipment
ASSISTANCE with operation
SUPPLY reagents and materials
METHOD DEVELOPMENT
SELECTION of methodology
EXPERIMENTAL design
DATA storage
METABOLICEXPERIMENTFLOWCHART
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Exp-1 Exp-2 Exp-3 Exp-4
ACCESS to the Equipment
ASSISTANCE with operation
SUPPLY reagents and materials
METHOD DEVELOPMENT
SELECTION of methodology
EXPERIMENTAL design
DATA storage
METABOLICEXPERIMENTFLOWCHART
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Exp-1 Exp-2 Exp-3 Exp-4
ACCESS to the Equipment
ASSISTANCE with operation
SUPPLY reagents and materials
METHOD DEVELOPMENT
SELECTION of methodology
EXPERIMENTAL design
DATA storage
METABOLICEXPERIMENTFLOWCHART
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Exp-1 Exp-2 Exp-3 Exp-4
ACCESS to the Equipment
ASSISTANCE with operation
SUPPLY reagents and materials
METHOD DEVELOPMENT
SELECTION of methodology
EXPERIMENTAL design
DATA storage
METABOLICEXPERIMENTFLOWCHART
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TECHNICALREPORT
REPORTS
DETAILEDSCIENTIFICREPORT
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Biologicalsamples:examplesfromongoingexperiments
Cancer cell lines (Leukemias, Lung adenocarcinoma,
Prostate and Cervical cancer..)
Normal eukaryotic cells (Fibroblasts, prostate cells
microglia)
Bacteria
Neurospheres (neural stem cells)
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DRUGS
TRANSFECTIONS
DIFFERENTIATION
STIMULATION WITH COMPOUNDS
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• STEP1:Accessisrequestedbye-mailtotheManagingdirectororadelegate(seecontacts).Experimentalbackground,numberofsamplestobeassayed….
• STEP2:EvaluationoftherequestbytheHypACBteam.Criteria:
Feasibilityoftheexperimentandtypeofaccessrequired.Scheduledpriority: Level1:Internalacademicusers. Level2:Externalacademicusers. Level3:Externalnon-academicinvestigators.
• STEP3:Experimentisrunbytheplatformpersonnel.
Theresultsoftheexperimentswillbeprovidedwithallexperimentaldetails[…].Rawdatawill be stored as a copy for 5 years, according to international rules for scientificcorrectness.Dataanalysiscanbeperformedbytheend-usersorbytheHypACBteambasedonspecificagreement.
USERPOLICIESANDGUIDELINES
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USERPOLICIESANDGUIDELINES
• STEP1:Accessisrequestedbye-mailtotheManagingdirectororadelegate(seecontacts).Experimentalbackground,numberofsamplestobeassayed….
• STEP2:EvaluationoftherequestbytheHypACBteam.Criteria:
Feasibilityoftheexperiment.Scheduledpriority: Level1:Internalacademicusers. Level2:Externalacademicusers. Level3:Externalnon-academicinvestigators.
• STEP3:Experimentisrun.TypeofaccessbytheHypACBpersonnel.
AcknowledgementsInvestigatorsarerequiredtoacknowledgetheHyp-ACBplatform(andSapienzaFinancialContributionbyGrantNumber)inanypublicationthatresultsfromtheuseoftheplatform.Namesofindividualsfromplatformwillbeincluded,ifpreviouslyagreedoriftheyprovidedanyintellectualinputoradditionaleffort.
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• STEP1:Accessisrequestedbye-mailtotheManagingdirectororadelegate(seecontacts).Experimentalbackground,numberofsamplestobeassayed…..
• STEP2:EvaluationoftherequestbytheHypACBteam.Criteria:
Feasibilityoftheexperimentandtypeofaccessrequired.Scheduledpriority: Level1:Internalacademicusers. Level2:Externalacademicusers. Level3:Externalnon-academicinvestigators.
• STEP3:TypeofaccessisagreedandExperimentisrun.
AcknowledgementsInvestigatorsarerequiredtoacknowledgetheHyp-ACBplatform(andSapienzaFinancialContributionbyGrantNumber)inanypublicationthatresultsfromtheuseoftheplatform.Namesofindividualsfromplatformwillbeincluded,ifpreviouslyagreedoriftheyprovidedanyintellectualinputoradditionaleffort.
USERPOLICIESANDGUIDELINES
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• STEP1:Accessisrequestedbye-mailtotheManagingdirectororadelegate(seecontacts).Experimentalbackground,numberofsamplestobeassayed……..
• STEP2:EvaluationoftherequestbytheHypACBteam.Criteria:
Feasibilityoftheexperiment.Scheduledpriority: Level1:Internalacademicusers. Level2:Externalacademicusers. Level3:Externalnon-academicinvestigators.
• STEP3:TypeofaccessisagreedandExperimentisrun.Resultsareprovidedandstored.
AcknowledgementsInvestigatorsarerequiredtoacknowledgetheHyp-ACBplatform(andSapienzaFinancialContributionby
GrantNumber)inanypublicationthatresultsfromtheuseoftheplatform.Namesofindividualsfromplatformwillbeincluded,ifpreviouslyagreedoriftheyprovidedany
intellectualinputoradditionaleffort.
USERPOLICIESANDGUIDELINES
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TYPEOFACCESS
Directaccessbyendusers
Experiment(fromcell
culturetodata)
Fullexperimentalplanningandfollow-up
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TimeallocationTime
MaterialsPersonnel
AnalysisInterpretation
Methoddevelopment
TYPEOFACCESS
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TimeallocationTime
MaterialsPersonnel
AnalysisInterpretation
Methoddevelopment
150€ 1200€
OFFICIAL PRICE LIST 2019 UNDER APPROVAL
+35%forhypoxia
ExampleofExperimentalCosts(Metabolism96-wellplate)
TYPEOFACCESS:Serviceonly
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FOR SAPIENZA USERS: COST REDUCTION IS PLANNED (APPROX. 30-50% )
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TimeallocationTime
MaterialsPersonnel
AnalysisInterpretation
Methoddevelopment
150€ 1200€
OFFICIAL PRICE LIST UNDER APPROVAL
+35%forhypoxia
ExampleofExperimentalCost(Metabolism96-wellplate)
TYPEOFACCESS:Collaboration
CHARGE REDUCTION
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Accessonly Experimentandaccess Full
TYPEOFACCESS:Training
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NEW REQUESTS WELLCOME!
TEMPORARY WEBSITE www.macinmec/hypacb-facility/
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CONTACTS
• Managing director: prof. Francesca Cutruzzolà [email protected] Tel. 0649910955
Platform Staff
Data Manager: Prof. Serena Rinaldo [email protected]
Cell Biology: Dr. Alessio Paone [email protected]
Technical staff: Dr. Giovanna Boumis [email protected]
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