同所膵臓癌マウスモデルのダイナミックコントラスト強化磁気共鳴画像
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
この方法の全体的な目的は、マウスにおける同所性膵臓腫瘍異種移植片のためのダイナミック造影磁気共鳴画像（DCE-MRI）を適用することである。 DCE-MRIは、注射後の一定期間にMRコントラストの変化を監視することにより、標的組織内の微小血管系を評価するための非侵襲的方法である。 DCE-MRIは、悪性腫瘍を診断し、種々の治療1-4腫瘍応答を評価するために利用されている。定量的DCE-MRIは、高い再現性5を提示した。標的組織におけるMR造影剤の薬物動態学的パラメータを定量するために、造影剤注入前の異なる時点およびT1マップで取得されたすべてのDCE-MR画像は、6を合わせが行われなければならない。しかし、腹部領域における呼吸器および蠕動運動に、定量的なDCE-MRIは、胃腸腫瘍の限定されたアプリケーションを持っています。
同所性膵臓腫瘍モデルを評価するために利用されている生物学的療法と化学療法7,8-以下の膵臓腫瘍応答。元の腫瘍部位での微小環境を反映し、それによって治療に対するヒト腫瘍の応答をより正確に予測することができるので、同所性腫瘍モデルは、従来の皮下モデルよりも優れて考えられている。しかし、マウスの膵臓は腹部の左上象限に位置しているので、マウスにおける同所性膵臓腫瘍異種移植片の定量的なDCE-MRIは、容易に実施されていない。
我々は、胸部9から運動伝達を防止するために直角に曲げられたプラスチック基板を使用して腫瘍を固定することにより、マウスにおける腹部腫瘍のDCE-MRIのプロトコールを確立した。このボードによって適用される圧力は腹部領域に局在していた、と呼吸困難をもたらしていない。自動画像coregistration技術は、自由呼吸モードの腹部臓器のDCE-MRIのために検証されているが、それはeffectivelを行い、yが唯一の標的領域は、ゆっくりと定期的に10移動したとき。動物の呼吸速度は、画像形成時に可変であるため、腹部領域における身体拘束は、同所性膵臓腫瘍のマウスモデルにおける信頼性の高い薬物動態パラメータを取得するために必要であろう。我々は、正常にDCE-MRI 11-13に直角に折り曲げられたプラスチック基板を用いた同所性膵臓腫瘍異種移植片におけるMR造影剤の薬物動態学的パラメータを定量化している。ここでは、同所性膵臓腫瘍モデルマウスにおける腫瘍異種移植片のDCE-MRI、および薬物動態学的パラメータの定量化の詳細な手順を提示する。
Protocol
Representative Results
Discussion
我々は、マウスにおいて、免疫不全マウスを用いて、腹部腫瘍のDCE-MRIを同所性膵臓腫瘍モデルの詳細な方法を導入し、その動力学的パラメータを定量化している。膵臓の尾に針を挿入するときに同所膵臓腫瘍モデリングでは、注意が必要です。成功した場合、細胞は小胞を作る膵臓の頭部に転送されます。直角に曲がったプラスチック基板を適用する場合、それは腫瘍が板の上端の下方に位…
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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