이 작품은 국립 보건 의학 연구위원회, 호주 (1,008,865), 호주 연구위원회 (LE110100125), 국립 암 연구소 (CA138687-01), 에리카 슬론은 국립 유방암의 초기 경력 원정대에 의해 지원됩니다에 의해 지원되었다 재단, 호주. 코리나 김 푹스는 스위스 암 리그와 제약 과학의 모나 쉬 대학에서 HDR 장학금의 친교에 의해 지원됩니다. Eliane의 불안은 베른 암 리그에서 교부금에 의해 지원됩니다.

증명되는 프로토콜은 저자의 기관의 동물 관리 및 사용위원회의지도 및 승인하에 수행됩니다. 모든 실험은 모든 관련 지침, 규정 및 규제 기관 준수에 실행됩니다. 1. 췌장암 세포 라인을 transducing에 <ol><li> 앞에서 설명한대로 형질 도입 췌장암 세포는 루시 페라 제 표현. 12,13 테드 스티븐스 - 1 Capan-1 췌장암 세포주 반딧불 루시 페라로 형질 여기에 사용됩니다. </li></ol> 참고 : Renilla의 루시 페라 나 세균 루시퍼도 사용할 수 있습니다. 2. 췌장암 세포의 준비 <ol><li> 문화가 70 % 합류 할 때까지 췌장암 세포를 형질. </li><li> 췌장 세포를 올리고 가능성 90 % 이상되어 있는지 확인합니다. </li><li> 냉장 마트 리젤의 3:2 혼합물에서 2 × 10 7 세포 / ML에 resuspend을 : 인산 염은 버퍼오프라인 (PBS). </li><li> 이전 췌장에 주입 얼음 마트 리젤 세포 현탁액을 유지합니다. </li></ol> 참고 : 마트 리젤의 빠른 응고을 보장하기 위해, 세포 펠렛의 양을 고려하여 PBS 볼륨을 줄일 수 있습니다. 이전 주입 응고를 방지하기 위해 항상 얼음처럼 차가운 악기와 주사기를 사용하여 마트 리젤을 처리합니다. 제안 된 휴대폰 번호는 가이드와 각 세포주에 대해 경험적으로 결정되어야한다. 3. 마우스 준비 <ol><li> 2-3% isoflurane을 흡입하여 마우스를 마취. 부드러운 발가락 핀치에 페달 반사의 부족에 의한 마취의 깊이를 결정합니다. </li><li> 건조를 방지하기 위해 눈에 윤활제를 바릅니다. </li><li> 37 ° C 가열 패드의 뒷면에 마우스를 놓고 조심스럽게 복부의 왼쪽을 높이기 위해 마우스를 켭니다. </li><li> 10 % 포비돈 요오드 용액으로 복부를 준비합니다. </li></ol> 참고 : Injectab르 마취 흡입 마취 대신 사용할 수 있습니다. 수술 전 금식이 필요하지 않습니다. 4. 개복술 <ol><li> 불임 수술기구를 사용하면 대략 1 센티미터 정중선에서 옆 왼쪽​​으로 피부에 1.5 cm의 절개를합니다. </li><li> 기본 복부 근육에 1.5 cm의 절개를합니다. </li><li> 집게를 사용하여 비장을 찾아 부드럽게 복강에서 비장을 제거합니다. 기본 췌장을 노출하는 멸균 면봉에 따라서 비장을 보호합니다. </li><li> 비장에 인접한 췌장의 꼬리를 찾습니다. </li><li> 29 G 0.3 ML 인슐린 주사기 사용, 췌장에 마트 리젤 세포 현탁액의 20 μl를 주입. </li><li> 마트 리젤이 응고 할 때까지 주입 후, 30 ~ 60 초 동안 췌장에 주사기를 누릅니다. 이 중요한 단계는 세포의 누출을 최소화합니다. </li><li> 누설이 발생하지 확인하기 위해 주사 부위를 검사합니다. </li><li> 복강에 비장과 췌장을 반환합니다. </li></ol> 참고 : 얇은 될 수있는 췌장의 등쪽면을 펑 쳐링 않도록주의하십시오. 5. 복부 벽 마감 <ol><li> 연속 스티치를 사용하여 원형 바늘로 흡수 꼰 4-0 봉합사와 마우스의 복부 근육을 닫습니다. </li><li> 연속 스티치를 사용하여 절단 바늘 비 흡수성 필라멘트 6-0 봉합사로 외부 피부를 닫습니다. </li><li> 흡입 마취제 및 피하 주입 0.05-0.1 ㎎ / ㎏ 부 프레 노르 핀에서 마우스를 제거합니다. </li><li> 마우스가 음식과 물을 무료로 이용할와 37 ° C 가열 패드에 배치의 새장에 복구 할 수 있습니다. 쥐 같은 hunching 또는 감소 이동성 고통의 흔적을 보여 경우, 부 프레 노르 핀은 36 시간에 걸쳐 매 12 시간을 부여 할 수 있습니다. </li><li> 상처 치유 (7-10 일) 후, 마우스를 마취와 외부 봉합을 제거합니다. </li></ol> 6. 췌장 Canc의 발광 추적어 진행 <ol><li> isoflurane을 흡입하여 마우스를 마취. </li><li> 꼬리 정맥을 통해 150 ㎎ / ㎏ D-루시페린을 주입. </li><li> 발광 생물 이미징 시스템에 마우스를 놓고 앞에서 설명한대로 백색 빛과 생물 발광 영상을 캡처합니다. 14,15가 </li><li> 흡입 마취에서 마우스를 제거하고 그것의 홈 케이지에 복구 할 수 있습니다. </li></ol> 참고 : 바이오 루미 네 센트 영상은 비 침습적이며, 종양의 성장 속도론을 조사하기 위해 정기적으로 실시 할 수 있습니다. 췌장 꼬리의 이미지 종양에 그것의 왼쪽에 마우스를두고하는 것이 중요합니다, 카메라를 향해 종양 점 때문에. 주파수 전에 FOV 12.5 비닝 (binning) 4와 함께 살고 이미징 4.3.1 소프트웨어를 실행하는 루미나 II 이미징 시스템 (퍼킨 - 엘머, 이전 캘리퍼스 생명 과학)를 사용하여 실험 끝점에 일주일에 세 번까지 증가로 우리는 일주일에 한 번 군데 F-스톱 1, 노출 1-60 초 (높은 경험치에 의해 결정픽셀의 채도없이 osure).

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저자는 그들이 더 경쟁 재정적 이익이 없다는 것을 선언합니다.

여기에서 우리는 췌장 종양 개발 및 진행의​​ 경도 평가를위한 동소 모델을 설명합니다. 기본 종양의 성장 속도론을 재현 할 수있다 (그림 3)과 새로운 안티 - 췌장암 치료에 대한 종양 반응의 분석 예 : 루시 페라 제 태그가 세포의 생물 발광 영상을 사용하여 비 침습적으로 모니터링 할 수 있습니다. 인간의 질병과 일치 모델은 췌장의 미세와 종양 세포의 상호 작용을 조사 ?...

췌장암 4-6 %의 5 년 생존율, 암 관련 사망의 네 번째 주요 원인이다. 환자의 1,2 단지 15 %는 수술을받을 수 질병 조기에 진단하고, 종양이 재발된다 &gt;에서 그 환자의 80 %가. 3,4 젬시 타빈은 췌장 선암의 치료에 사용된다, 그러나, chemoresistance는 일반적이고 자주 약물 생존율에 거의 영향을 미친다. 췌장암을 치료하는 새로운 5 약리 전략이 매우 필요합니다. 그들의 개발은 치료 적 개입에 민감 할 수 있습니다 질병의 진행의 주요 단계를 크게 향상 이해에 따라 달라집니다. 췌장암의 동소 모델은 췌장암의 생물학을 공부 그들을 이상적인 도구를 만드는 인간의 질병의 주요 측면을 모방. 췌장암 세포 행동의 체외에서 세포 기반 분석과는 달리 6-9췌장암의 생체 내 모델에서 D 피하는 동소 모델은 췌장의 미세와 종양 세포의 상호 작용을 조사 할 수 있습니다. 질병 진행의 반응 속도는 동소 모델에서 높은 재현성하고 잘 새로운 치료제의 전임상 시험에 적합하게 짧은 시간 프레임 (주)에 발생합니다. 이 질병의 발병이 더 많은 변수 기간 (개월 1 년) 10. 더 공격적인 세포 라인과 함께 사용을 통해 발생 형질 전환 모델 대조적입니다, 췌장암의 동소 모델에서 본 것과 유사한 자연 전이의 패턴을 가지고 환자는. 같은 반딧불 루시 페라로 발광 리포터 유전자의 8 발현 종양 성장의 종 모니터링, 전이성 보급, 재발 및 치료에 대한 반응을 용이하게합니다. 6,11 여기에 우리가 MATR을 활용 췌장암의 동소 모델을 설명종양 진행의 비 침습 모니터링을위한 지역화 된 세포 전달 및 생체 내 생물 발광 이미징에 대한 이젤. 췌장암이 동소 모델은 질병의 진행 및 syngenic 또는 이종 이식 모델에서 치료 개입에 대한 응답의 비 침습 분석을 할 수 있습니다.

<table border="1">
 <tbody>
 <tr>
 <td>Equipment</td>
 <td>Company</td>
 <td>Catalog Number</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Clean Bench coat</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Heating pad</td>
 <td></td>
 <td></td>
 <td>Set to 37 &deg;C </td>
 </tr>
 <tr>
 <td>Ivis Lumina ll Bioluminescent imager</td>
 <td>Caliper </td>
 <td></td>
 <td>Alternative bioluminescent imaging systems include In vivo F PRO (Carestream) and Photon Imager (Biospace Lab)</td>
 </tr>
 <tr>
 <td>Dissecting scissors</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Iris forceps (serrated)</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Needle holder</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>27 G 0.3 ml insulin syringe</td>
 <td>Terumo</td>
 <td>T35525M2913</td>
 <td></td>
 </tr>
 </tbody>
</table>

bioluminescent orthotopic model of pancreatic cancer progression

<p class="jove_content"&gt; 췌장암 4-6 %의 매우 가난한 5 년 생존율이 있습니다. 새로운 치료 옵션이 매우 필요하고 췌장암 생물학의 이해 증진에 의존하고 있습니다. 더 나은 췌장의 미세과 암 세포의 상호 작용을 이해하기 위해, 우리는 암 진행의 비 침습 모니터링을 허용 췌장암의 동소 모델을 보여줍니다. 루시 페라 제 태그가 췌장암 세포는 리겔에 재현 탁하고 개복 수술 중에 췌장의 꼬리에 전달됩니다. Matrigel을 주입하는 동안 암 세포의 누출을 방지하기 위해 신체 온도에서 응고. 먼 장기에 기본 종양의 성장과 전이를 사용하여 루시 페라 제 기판 루시페린의 주입에 따라 모니터링됩니다<em&gt; 생체 내</em암 세포 생물 발광 방출의&gt; 영상.<em&gt; 생체 내</em&gt; 영상 또한 절제 후 일차 종양의 재발을 추적하는 데 사용할 수 있습니다. 이 동소 모델은 동계 및 이종 이식 모델 모두에 적합하고 기본 췌장 종양 전이의 성장 소설 안티 - 암 치료의 영향을 조사하기 위해 사전 임상 실험에 사용할 수 있습니다.</p&gt;

이 방법은 마취 유도, 개복술, 마트 리젤 및 복부 폐쇄 (그림 1A)에서 암 세포의 주입 등의 수술 절차를 사용하여 췌장암의 동소 모델을 설명합니다. 주입 된 세포는 췌장의 표면에 거품 (그림 1B)를 형성한다. 췌장암의 진행은 비 침습적 암 세포 증식 및 보급 (그림 2)를 추적하는 생체 생물 발광 이미징에 사용하여 모니터링 할 수 있습니다. 간 전?...

Watch this Scientific Journal Video about Bioluminescent Orthotopic Model of Pancreatic Cancer Progression at JoVE.com

Bioluminescent Orthotopic Model of Pancreatic Cancer Progression

췌장암 생물학의 개선 이해는 매우 췌장암을 치료하는 더 나은 치료 옵션의 개발을 가능하게하기 위해 필요합니다. 이러한 요구를 해결하기 위해, 우리는 사용하여 암 진행의 비 침습 모니터링을 허용 췌장암의 동소 모델을 보여<em&gt; 생체 내</em&gt; 생물 발광 영상.

췌장암의 진행 발광 동소 모델

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%.  New therapeutic options are critically needed and depend on improved ...

bioluminescent-orthotopic-model-of-pancreatic-cancer-progression

Research

JoVE Journal

Medicine

26.7K 조회수.  Monash University. 췌장암 생물학의 개선 이해는 매우 췌장암을 치료하는 더 나은 치료 옵션의 개발을 가능하게하기 위해 필요합니다. 이러한 요구를 해결하기 위해, 우리는 사용하여 암 진행의 비 침습 모니터링을 허용 췌장암의 동소 모델을 보여 생체 내 생물 발광 영상.

비디오: Bioluminescent Orthotopic Model of Pancreatic Cancer Progression

Murine Bioluminescent Hepatic Tumour Model

This video describes the establishment of liver metastases in a mouse model that can be subsequently analysed by bioluminescent imaging. Tumour cells are administered specifically to the liver to induce a localised liver tumour, via mobilisation of the spleen and splitting into two, leaving intact the vascular pedicle for each half of the spleen. Lewis lung carcinoma cells that constitutively express the firefly luciferase gene (luc1) are inoculated into one hemi-spleen which is then resected 10 minutes later. The other hemi-spleen is left intact and returned to the abdomen. Liver tumour growth can be monitored by bioluminescence imaging using the IVIS whole body imaging system. Quantitative imaging of tumour growth using IVIS provides precise quantitation of viable tumour cells. Tumour cell death and necrosis due to drug treatment is indicated early by a reduction in the bioluminescent signal. This mouse model allows for investigating the mechanisms underlying metastatic tumour-cell survival and growth and can be used for the evaluation of therapeutics of liver metastasis. 

This article describes a procedure for the induction of orthotopic bioluminescent liver tumours in mice, and subsequent analysis of tumour growth confined to the liver using live whole body luminescence imaging.

Murine 발광 생물 간장 Tumour 모델

This video describes the establishment of liver metastases in a mouse model that can be subsequently analysed by bioluminescent imaging. Tumour cells are ...

연구

의학

An Orthotopic Model of Murine Bladder Cancer

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and subsequent cell adhesion. After their bladders are transurethrally catheterized and drained, animals are again catheterized to permit insertion of a platinum wire into bladders without damaging the urethra or bladder. The catheters are made of Teflon to serve as an insulator for the wire, which will conduct electrical current into the bladder to create a burn injury. An electrocautery unit is used to deliver 2.5W to the exposed end of the wire, burning away extracellular layers and providing attachment sites for carcinoma cells that are delivered in suspension to the bladder through a subsequent catheterization. Cells remain in the bladder for 90 minutes, after which the catheters are removed and the bladders allowed to drain naturally. The development of tumor is monitored via ultrasound. Specific attention is paid to the catheterization technique in the accompanying video.

Bladder tumors are established in female mice in a minimally invasive fashion through catheterization, local cauterization, and subsequent adhesion of carcinoma cells to the burn sites.

Murine 방광 암의 Orthotopic 모델

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and ...

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient management strategies. Orthotopic mouse models of oral cancer have been developed to facilitate the study of factors that impact invasion and serve as model system for evaluating anti-tumor therapeutics. In these systems, visualization of disseminated tumor cells within oral cavity tissues has typically been conducted by either conventional histology or with in vivo bioluminescent methods. A primary drawback of these techniques is the inherent inability to accurately visualize and quantify early tumor cell invasion arising from the primary site in three dimensions. Here we describe a protocol that combines an established model for squamous cell carcinoma of the tongue (SCOT) with two-photon imaging to allow multi-vectorial visualization of lingual tumor spread. The OSC-19 head and neck tumor cell line was stably engineered to express the F-actin binding peptide LifeAct fused to the mCherry fluorescent protein (LifeAct-mCherry). Fox1nu/nu mice injected with these cells reliably form tumors that allow the tongue to be visualized by ex-vivo application of two-photon microscopy. This technique allows for the orthotopic visualization of the tumor mass and locally invading cells in excised tongues without disruption of the regional tumor microenvironment. In addition, this system allows for the quantification of tumor cell invasion by calculating distances that invaded cells move from the primary tumor site. Overall this procedure provides an enhanced model system for analyzing factors that contribute to SCOT invasion and therapeutic treatments tailored to prevent local invasion and distant metastatic spread. This method also has the potential to be ultimately combined with other imaging modalities in an in vivo setting.

A comprehensive overview of the techniques involved in generating a mouse model of oral cancer and quantitative monitoring of tumor invasion within the tongue through multi-photon microscopy of labeled cells is presented. This system can serve as a useful platform for the molecular assessment and drug efficacy of anti-invasive compounds.

구강 편평 세포 암종의 Orthotopic 마우스 모델에서 종양 세포 침략의 다중 광자 이미징

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

In vivo Dual Substrate Bioluminescent Imaging

Our understanding of how and when breast cancer cells transit from established primary tumors to metastatic sites has increased at an exceptional rate since the advent of in vivo bioluminescent imaging technologies 1-3. Indeed, the ability to locate and quantify tumor growth longitudinally in a single cohort of animals to completion of the study as opposed to sacrificing individual groups of animals at specific assay times has revolutionized how researchers investigate breast cancer metastasis. Unfortunately, current methodologies preclude the real-time assessment of critical changes that transpire in cell signaling systems as breast cancer cells (i) evolve within primary tumors, (ii) disseminate throughout the body, and (iii) reinitiate proliferative programs at sites of a metastatic lesion. However, recent advancements in bioluminescent imaging now make it possible to simultaneously quantify specific spatiotemporal changes in gene expression as a function of tumor development and metastatic progression via the use of dual substrate luminescence reactions. To do so, researchers take advantage for two light-producing luciferase enzymes isolated from the firefly (Photinus pyralis) and sea pansy (Renilla reniformis), both of which react to mutually exclusive substrates that previously facilitated their wide-spread use in in vitro cell-based reporter gene assays 4. Here we demonstrate the in vivo utility of these two enzymes such that one luminescence reaction specifically marks the size and location of a developing tumor, while the second luminescent reaction serves as a means to visualize the activation status of specific signaling systems during distinct stages of tumor and metastasis development. Thus, the objectives of this study are two-fold. First, we will describe the steps necessary to construct dual bioluminescent reporter cell lines, as well as those needed to facilitate their use in visualizing the spatiotemporal regulation of gene expression during specific steps of the metastatic cascade. Using the 4T1 model of breast cancer metastasis, we show that the in vivo activity of a synthetic Smad Binding Element (SBE) promoter was decreased dramatically in pulmonary metastasis as compared to that measured in the primary tumor 4-6. Recently, breast cancer metastasis was shown to be regulated by changes within the primary tumor microenvironment and reactive stroma, including those occurring in fibroblasts and infiltrating immune cells 7-9. Thus, our second objective will be to demonstrate the utility of dual bioluminescent techniques in monitoring the growth and localization of two unique cell populations harbored within a single animal during breast cancer growth and metastasis.

In vivo Dual Substrate Bioluminescent Imaging

Herein we describe the methods to construct, visualize, and quantify the bioluminescent reactions of both firefly and renilla luciferase enzymes expressed in metastatic breast cancer cells during their growth and metastasis in vivo.

생체 듀얼 기판 발광 생물 이미징에

Our understanding of how and when breast cancer cells transit from established primary tumors to metastatic sites has increased at an exceptional rate ...

Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model

Glioblastoma multiforme (GBM) are the most common and aggressive adult primary brain tumors1. In recent years there has been substantial progress in the understanding of the mechanics of tumor invasion, and direct intracerebral inoculation of tumor provides the opportunity of observing the invasive process in a physiologically appropriate environment2. As far as human brain tumors are concerned, the orthotopic models currently available are established either by stereotaxic injection of cell suspensions or implantation of a solid piece of tumor through a complicated craniotomy procedure3. In our technique we harvest cells from tissue culture to create a cell suspension used to implant directly into the brain. The duration of the surgery is approximately 30 minutes, and as the mouse needs to be in a constant surgical plane, an injectable anesthetic is used. The mouse is placed in a stereotaxic jig made by Stoetling (figure 1). After the surgical area is cleaned and prepared, an incision is made; and the bregma is located to determine the location of the craniotomy. The location of the craniotomy is 2 mm to the right and 1 mm rostral to the bregma. The depth is 3 mm from the surface of the skull, and cells are injected at a rate of 2 &mu;l every 2 minutes. The skin is sutured with 5-0 PDS, and the mouse is allowed to wake up on a heating pad. From our experience, depending on the cell line, treatment can take place from 7-10 days after surgery. Drug delivery is dependent on the drug composition. For radiation treatment the mice are anesthetized, and put into a custom made jig. Lead covers the mouse's body and exposes only the brain of the mouse. The study of tumorigenesis and the evaluation of new therapies for GBM require accurate and reproducible brain tumor animal models. Thus we use this orthotopic brain model to study the interaction of the microenvironment of the brain and the tumor, to test the effectiveness of different therapeutic agents with and without radiation.

The purpose of this article is to describe the use of an orthotopic glioblastoma model for chemoradiation studies. This article will go though cell processing, implanting, and radiotherapy of the mouse using an intracranial model.

Orthotopic 마우스 뇌종양 모델의 조합 방사선 치료

Glioblastoma multiforme (GBM) are the most common and aggressive adult primary brain tumors1. In recent years there has been substantial progress in the ...

An Orthotopic Bladder Tumor Model and the Evaluation of Intravesical saRNA Treatment

We present a novel method for treating bladder cancer with intravesically delivered small activating RNA (saRNA) in an orthotopic xenograft mouse bladder tumor model. The mouse model is established by urethral catheterization under inhaled general anesthetic. Chemical burn is then introduced to the bladder mucosa using intravesical silver nitrate solution to disrupt the bladder glycosaminoglycan layer and allows cells to attach. Following several washes with sterile water, human bladder cancer KU-7-luc2-GFP cells are instilled through the catheter into the bladder to dwell for 2 hours. Subsequent growth of bladder tumors is confirmed and monitored by in vivo bladder ultrasound and bioluminescent imaging. The tumors are then treated intravesically with saRNA formulated in lipid nanoparticles (LNPs). Tumor growth is monitored with ultrasound and bioluminescence. All steps of this procedure are demonstrated in the accompanying video.

Establishing an orthotopic bladder tumor model to evaluate antitumor effects of intravesically delivered saRNA and monitoring tumor growth by ultrasound and bioluminescent imaging.

Orthotopic 방광 종양 모델 및 Intravesical saRNA 트리 트먼트의 평가

We present a novel method for treating bladder cancer with intravesically delivered small activating RNA (saRNA) in an orthotopic xenograft mouse bladder ...

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as studies on graft preservation and ischemia-reperfusion injury 1,2, immunological responses 3,4, hemodynamics 5,6, and small-for-size syndrome 7. The rat OLT is among the most difficult animal models in experimental surgery and demands advanced microsurgical skills that take a long time to learn. Consequently, the use of this model has been limited. Since the reliability and reproducibility of results are key components of the experiments in which such complex animal models are used, it is essential for surgeons who are involved in rat OLT to be trained in well-standardized and sophisticated procedures for this model.
While various techniques and modifications of OLT in rats have been reported 8 since the first model was described by Lee et al. 9 in 1973, the elimination of the hepatic arterial reconstruction 10 and the introduction of the cuff anastomosis technique by Kamada et al. 11 were a major advancement in this model, because they simplified the reconstruction procedures to a great degree. In the model by Kamada et al., the hepatic rearterialization was also eliminated. Since rats could survive without hepatic arterial flow after liver transplantation, there was considerable controversy over the value of hepatic arterialization. However, the physiological superiority of the arterialized model has been increasingly acknowledged, especially in terms of preserving the bile duct system 8,12 and the liver integrity 8,13,14.
In this article, we present detailed surgical procedures for a rat model of OLT with hepatic arterial reconstruction using a 50% partial graft after ex vivo liver resection. The reconstruction procedures for each vessel and the bile duct are performed by the following methods: a 7-0 polypropylene continuous suture for the supra- and infrahepatic vena cava; a cuff technique for the portal vein; and a stent technique for the hepatic artery and the bile duct.

Orthotopic liver transplantation in rats is an indispensable experimental model for biomedical research. Here we present our surgical procedures for orthotopic rat liver transplantation with hepatic arterial reconstruction using a 50% partial graft.

간 간선 재건과 부분 Orthotopic 간 이식의 쥐 모델에 대한 외과 절차

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

An Orthotopic Bladder Cancer Model for Gene Delivery Studies

Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are needed. The tumor microenvironment can significantly influence tumor development and therapy response. It is therefore often desirable to grow tumor cells in the organ from which they originated. This protocol describes an orthotopic model of bladder cancer, in which MB49 murine bladder carcinoma cells are instilled into the bladder via catheterization. Successful tumor cell implantation in this model requires disruption of the protective glycosaminoglycan layer, which can be accomplished by physical or chemical means. In our protocol the bladder is treated with trypsin prior to cell instillation. Catheterization of the bladder can also be used to deliver therapeutics once the tumors are established. This protocol describes the delivery of an adenoviral construct that expresses a luciferase reporter gene. While our protocol has been optimized for short-term studies and focuses on gene delivery, the methodology of mouse bladder catheterization has broad applications.

Implantation of cancer cells into the organ of origin can serve as a useful preclinical model to evaluate novel therapies. MB49 bladder carcinoma cells can be grown within the bladder following intravesical instillation. This protocol demonstrates catheterization of the mouse bladder for the purpose of tumor implantation and adenoviral delivery.

유전자 전달 연구 동소 방광암 모델

Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are ...

An Orthotopic Murine Model of Human Prostate Cancer Metastasis

Our laboratory has developed a novel orthotopic implantation model of human prostate cancer (PCa). As PCa death is not due to the primary tumor, but rather the formation of distinct metastasis, the ability to effectively model this progression pre-clinically is of high value. In this model, cells are directly implanted into the ventral lobe of the prostate in Balb/c athymic mice, and allowed to progress for 4-6 weeks. At experiment termination, several distinct endpoints can be measured, such as size and molecular characterization of the primary tumor, the presence and quantification of circulating tumor cells in the blood and bone marrow, and formation of metastasis to the lung. In addition to a variety of endpoints, this model provides a picture of a cells ability to invade and escape the primary organ, enter and survive in the circulatory system, and implant and grow in a secondary site. This model has been used effectively to measure metastatic response to both changes in protein expression as well as to response to small molecule therapeutics, in a short turnaround time.

This orthotopic model of human prostate cancer allows for quantification of tumor size, circulating tumor cells, and formation of distinct metastasis to the lung. As cells must escape the primary organ, enter the blood stream, and implant into a secondary site, this model effectively recapitulates the scenario in humans.

인간의 전립선 암 전이의 동소 쥐과 모델

Our laboratory has developed a novel orthotopic implantation model of  human prostate cancer (PCa). As PCa death is not due to the primary tumor, but  ...

Dynamic Contrast Enhanced Magnetic Resonance Imaging of an Orthotopic Pancreatic Cancer Mouse Model

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.

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.