Dynamisk kontrast Forbedret Magnetic Resonance Imaging af en ortotopisk kræft i bugspytkirtlen Mouse Model
Summary
The goal of this protocol is to apply dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) for orthotopic pancreatic tumor xenografts in mice. DCE-MRI is a non-invasive method to analyze microvasculature in a target tissue, and useful to assess vascular response in a tumor following a novel therapy.
Abstract
Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) has been limitedly used for orthotopic pancreatic tumor xenografts due to severe respiratory motion artifact in the abdominal area. Orthotopic tumor models offer advantages over subcutaneous ones, because those can reflect the primary tumor microenvironment affecting blood supply, neovascularization, and tumor cell invasion. We have recently established a protocol of DCE-MRI of orthotopic pancreatic tumor xenografts in mouse models by securing tumors with an orthogonally bent plastic board to prevent motion transfer from the chest region during imaging. The pressure by this board was localized on the abdominal area, and has not resulted in respiratory difficulty of the animals. This article demonstrates the detailed procedure of orthotopic pancreatic tumor modeling using small animals and DCE-MRI of the tumor xenografts. Quantification method of pharmacokinetic parameters in DCE-MRI is also introduced. The procedure described in this article will assist investigators to apply DCE-MRI for orthotopic gastrointestinal cancer mouse models.
Introduction
Det overordnede mål med denne metode er at anvende dynamisk kontrast forbedret magnetisk resonans (DCE-MRI) for ortotopisk pancreas tumorxenotransplantater i mus. DCE-MRI er en ikke-invasiv metode til at vurdere mikrovaskulaturen i et målvæv ved at overvåge ændringen i MR kontrast over en vis periode efter injektion. DCE-MRI er blevet anvendt til at diagnosticere maligne tumorer og vurdere tumorrespons til forskellige terapier 1-4. Kvantitativ DCE-MRI har fremlagt høj reproducerbarhed 5. At kvantificere farmakokinetiske parametre for et MR-kontrastmiddel i et målvæv skal alle DCE-MR-billeder erhvervet på forskellige tidspunkter og T1 kort opnået før kontrast injektion coregistered 6. Men på grund af respiratoriske og peristaltiske bevægelser i maveregionen, er kvantitativ DCE-MRI havde begrænset ansøgning om gastrointestinale tumorer.
Orthotopisk pancreas tumormodeller er blevet anvendt til at vurderepancreas-tumor respons efter biologiske terapier og kemoterapier 7,8. Ortotopisk tumormodeller anses overlegen konventionelle subkutane modeller, da mikromiljøet i den oprindelige tumorsted reflekteres og derved human tumor respons på behandlingen kan mere nøjagtigt forudsiges. Imidlertid er muse pancreas placeret i den øverste venstre kvadrant af maven, så kvantitativ DCE-MRI af ortotopisk pancreas tumorxenotransplantater hos mus er ikke let gennemføres.
Vi har etableret en protokol af DCE-MRI af abdominale tumorer hos mus ved fastsættelse af tumorer ved hjælp af en retvinklet bøjet plastik bord for at forhindre bevægelse overførsel fra brystregionen 9. Det pres, som dette forum blev lokaliseret på maveregionen og har ikke medført åndedrætsbesvær. En automatiseret billede coregistration teknik er blevet godkendt til DCE-MRI af abdominale organer i en fri vejrtrækning tilstand, men det udfører effectively, når målregionerne bevæger sig langsomt og regelmæssigt 10. Respirationsfrekvens af dyr er variabel under billedbehandling, vil være nødvendigt så fysisk tilbageholdenhed i maveregionen at hente pålidelige farmakokinetiske parametre i ortotopisk pancreas tumor musemodeller. Vi har med succes kvantificeret de farmakokinetiske parametre af et MR-kontrastmiddel i ortotopisk pancreas tumorxenotransplantater hjælp af ortogonalt bøjet plast bord i DCE-MRI 11-13. Her præsenterer vi den detaljerede procedure for orthotopisk pancreas tumor modellering, DCE-MRI af tumorxenografter i mus og kvantificering af farmakokinetiske parametre.
Protocol
Representative Results
Discussion
Vi har indført de detaljerede metoder til orthotopisk pancreas tumor modellering ved hjælp af immunsvækkede mus, DCE-MRI af abdominale tumorer i mus og kvantificering af dets kinetiske parametre. I orthotopisk pancreas tumor modellering, skal man passe på, når du indsætter en nål i halen af bugspytkirtlen. Hvis det lykkes, vil cellerne blive overført til lederen af bugspytkirtlen skabe en lille bleb. Ved anvendelse af en retvinklet bøjet plast bord, er det vigtigt at bekræfte, at svulsten er placer…
Disclosures
The authors have nothing to disclose.
Acknowledgements
Authors thank Jeffrey Sellers to assist orthotopic pancreatic cancer mouse modeling. This work was supported by Research Initiative Pilot Awards from the Department of Radiology at UAB and NIH grants 2P30CA013148 and P50CA101955.
Materials
| Name of Material/ Equipment | Company | Catalog Number | Comments/Description |
| DMEM | Invitrogen | 11965-118 | |
| Fetal bovine serum | Harlan Laboratories | BT-9501 | |
| Betadine | Purdue products | 67618-153-01 | |
| 5-0 Prolene sutures | Ethicon | 8720H | |
| 9.4T MR scanner | Bruker Biospin Corporation | BioSpec 94/20 USR | |
| Gadoteridol | Bracco Diagnostics Inc | NDC 0270-1111-03 | |
| Micro-polyethelene tube | Strategic Applications, Inc | #PE-10-25 | |
| 30G blunt tip needle | Strategic Applications, Inc | 89134-194 | |
| Monitoring and gating system | SA instruments, Inc | Model 1030 | This is an MR compatiable system to measure resiratory rating and body temperature of small animals at the same time. |
| Syringe pump | New Era Pump Systems, Inc. | NE-1600 |
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