Dynamic Contrast Forbedret Magnetic Resonance Imaging av en ortotopiske Bukspyttkjertelkreft 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 målet med denne metoden er å bruke dynamisk kontrastforsterket magnetisk resonanstomografi (DCE-MRI) for ortotopiske bukspyttkjertel tumorxenotransplantater hos mus. DCE-MRI er en ikke-invasiv metode for å vurdere mikrovaskulaturen i et målvev ved å overvåke endringen av MR-kontrast over en viss tid etter injeksjon. DCE-MRI har vært benyttet til å diagnostisere ondartede svulster og å vurdere tumorrespons på ulike behandlinger 1-4. Kvantitative DCE-MRI har presentert høy reproduserbarhet 5. For å kvantifisere farmakokinetiske parameterne til en MR-kontrastmiddel i et mål vev, må alle DCE-MR-bilder ervervet ved ulike tidspunkt og T1 kart innhentes før kontrastinjeksjon bli coregistered 6. Men på grunn av luftveis og peristaltiske bevegelser i mageområdet, har kvantitative DCE-MRI hadde begrenset anvendelse for gastrointestinale svulster.
Ortotopisk pankreatiske tumormodeller er blitt benyttet for å vurderepankreas-tumor respons etter biologiske terapier og chemotherapies 7,8. Ortotopiske tumormodeller anses overlegen til konvensjonell subkutane modeller, siden mikromiljøet i den opprinnelige svulsten nettstedet er reflektert og dermed menneskelig tumor respons på behandlingen kan bli mer nøyaktig spådde. Imidlertid er muse bukspyttkjertelen ligger i den venstre øvre kvadrant av abdomen, så kvantitativ DCE-MRI of ortotopiske pankreas tumorxenografter i mus har ikke vært lett realiseres.
Vi har etablert en protokoll av DCE-MRI av magesvulster hos mus ved å feste svulster ved hjelp av en vinkelrett bøyd plast bord for å forhindre bevegelse overføring fra brystet regionen ni. Trykket brukes av dette forumet ble lokalisert på mageregionen, og har ikke resultert i pustevanskeligheter. En automatisert bilde coregistration teknikken har blitt validert for DCE-MRI av abdominal organer i en gratis-pust-modus, men det utfører effectively bare når målregioner bevege seg sakte og regelmessig 10. Respirasjonsfrekvens av dyr er variabel under avbildning, vil så fysisk tilbakeholdenhet i mageområdet være nødvendig å hente pålitelige farmakokinetiske parametere i ortotopiske bukspyttkjertelen svulst musemodeller. Vi har med hell kvantifisert de farmakokinetiske parameterne til en MR-kontrastmiddel i ortotopiske bukspyttkjertelen tumorxenotransplantater bruker orthogonally bøyd plast bord i DCE-MRI 11-13. Her presenterer vi detaljert prosedyre for orthotopic bukspyttkjertelen svulst modellering, DCE-MRI av tumorxenotransplantater hos mus, og kvantifisering av farmakokinetiske parametre.
Protocol
Representative Results
Discussion
Vi har innført de detaljerte metoder for orthotopic bukspyttkjertelen svulst modellering ved hjelp av immunsvikt mus, DCE-MRI av magesvulster hos mus, og kvantifisering av sine kinetiske parametre. I orthotopic bukspyttkjertelen svulst modellering, må det utvises forsiktighet når du setter inn en nål inn i halen av bukspyttkjertelen. Hvis de lykkes, vil cellene bli overført til leder av bukspyttkjertelen lage en liten bleb. Når du søker en orthogonally bøyd plast bord, er det avgjørende å bekrefte at svulsten …
Divulgations
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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