Name: a "patient-like" orthotopic syngeneic mouse model of hepatocellular carcinoma metastasis
Uploaded: 2015-10-24T14:00:00.000+00:00
Duration: 5 min 6 s
Description: Watch this Scientific Journal Video about A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis at JoVE.com

We would like to thank Pascal DuBois for proof-reading the article. The authors would also like to thank the personnel of the Animal Facility at Hunter College of The City University of New York. This work was supported by a NIH RCMI grant to Hunter College.

윤리 선언문 모든 동물 실험은 뉴욕 시립 대학의 헌터 대학의 기관 동물 관리 및 사용위원회 (IACUC)에 의해 승인되었다. 주 : 에이트 BALB / c 마우스는이 실험 절차에서 사용 하였다. 다섯 BALB / c 마우스는 차 종양을 생성하기 위해 5 X 106 BNL 1ME A.7R.1 마우스 간암 세포를 이식 하였다. 아니오 주입 세 BALB / c 마우스는 대조군으로 사용 하였다. BNL 1ME A.7R.1은 쥐의 비 종양 간세포 세포주 BNL c12에에서 화학 변환에 의해 유도 된 쥐의 간세포 암 세포주이다. BNL 1ME A.7R.1는 아메리칸 타입 컬쳐 콜렉션 (ATCC)로부터 수득 하였다. 1. 사전 수술 <ol><li> 모든 생존 수술은 건강을 위해 그 쥐를 확인합니다. 마우스가 건강하도록하려면, 피부의 텐팅 (tenting)에 의해 탈수의 징후를 검사합니다. 질병 또는 d의 증거와 마우스를 사용하지 마십시오이 실험에서 istress. </li><li> 그들은 저체중하지 보장하기 위해 마우스의 무게를. 무게가 적어도 16-20g 있는지 확인하십시오. </li><li> 마우스 당 주입 5 × 10 6 BNL 1ME A.7R.1 마우스의 간세포 암 세포를 준비합니다 <ol><li> BNL 1ME A.7R.1 마우스 간암 세포의 70 % 융합 성 세포 배양 접시 - 10 ml의 60 %에서 대기음 매체. 5 ml의 PBS로 두 번 씻는다. 접시에 트립신 2 ML을 추가하고 5 분 동안 공기와 5 % CO 2 가습 배양기에서 37 ° C에서 품어. </li><li> 그 후, 피펫 10 μL은 10 배의 배율 현미경으로 세포 계수에 대한 수동 혈구로 세포를 트립신. 4 개의 큰 코너에 위치한 사각형 셀들의 총 수를 카운트하고 세포의 총 수에 의해 희석 계수를 곱한다. 이어서, (ML 당 셀) 카운트 제곱의 수로 나누어 세포 농도 10 (6)에 의해 곱. </li><li> 계산을 이용하여, 무균 마우스 당 500 만 세포를 수집1.5 ml의 마이크로 원심 튜브. 1.5 ml의 마이크로 원심 튜브 레이블. </li></ol></li><li> 포스페이트 완충 염수 100 ㎕ (PBS)에 5 × 106 BNL 1ME A.7R.1 마우스 HCC 세포 재현 탁. 수술에서 이식까지 얼음이 세포를 보관하십시오. </li><li> 마취의 유도 및 머리 오염을 방지하기 위해 멸균 장비와 물질을 멀리 클리핑 등의 수술에 대한 쥐의 준비에 필요한 활동을 수행합니다. </li><li> 분당 1 L (약. 2.1 %)의 농도를 일정한 유량으로 산소와 함께 기화기 이소 플루 란을 사용하여 이소 플루 란 흡입에 의해 마우스의 전신 마취를 유도한다. 주 : 시스템은 자립이며, 정밀 기화기, 산소 공급과 가스 / 마취제 소기 필요한 호흡 회로에 연결된 캐니스터 목탄을 포함한다. </li><li> 마취는 수술 중 통증을 겪고에서 동물을 방지하기 위해 적절한 지 확인하십시오. 발가락 핀치를 수행하고 비 respon 위해 동물을 관찰siveness 마취를 확인합니다. </li><li> 이 때, 코 콘에 밸브 라인, 인감 상자 열기를 전환합니다. </li><li> 즉시 마우스를 마취로, 건조 및 손상 예방 조치로 각막에 안과 연고의 작은 금액을 적용합니다. </li><li> 이어서, 노즈콘 제대로 (유지 마취)에 배치하고 연결하여, 준비 공간에 동물을 이동. 이 준비 영역에서 탈모에 의해 수술을위한 수술 절개 부위를 준비합니다. </li><li> 수술의 영역이 다른 모든 활동이없는 위치에 있는지 확인합니다. </li><li> 작업 표면이 견고하고 적절하게 소독, unsoiled과 혼란이없는 수술 전인지 확인하십시오. 리넨 또는 이상 수술 트레이의 살균 조각을 놓습니다. </li><li> 수술 필드는 수술을 수행하는 사람 만 사용할 수 있는지 확인합니다. 수술 방 적절한 소독과 청소하고 적절한 세제를 휩쓸 및 분사되어 있는지 확인ctant. </li><li> 그것은 또한 거리에 문 및 공기 흐름 등의 창에서 위치 사이트의 오염에 이르는 먼지를 확산 할 수 있는지 확인합니다. </li><li> 멸균 악기 생존 수술을 수행합니다. 어느 제조업체의 권장하거나 오토 클레이브에 따라 살균제에 악기를 흡수. 사용할 준비가 기본 멸균 공급 장치를 사용합니다. </li><li> 별도의 장비는 기능에 따라 멸균 지역의 오염을 방지하기 위해. </li><li> 동물의 간을 액세스 할 수 절개 (중간 선 절개)의 사이트 주위에 약 1 인치를 면도. 수술의 사이트에서 멀리 동물에서 머리를 제거합니다. </li><li> 면도 후, 비누 살균제 (betadine 수술 스크럽)와 영역을 문질러 멸균 PBS 용액에 과량의 소독제를 씻어. 약 5 분 동안 지속 전체 스크럽 과정이 여러 번 수행합니다. 제거하고 사용 장갑을 폐기. </li><li> 그 후, 같은 surgic로 보호 복으로 변경알 가운, 스크럽 상단, 얼굴 마스크입니다. 다음으로, 수술 장갑 착용하기 전에 적절한 스크럽 솔루션 손을 소독. 참고 :이 흘리기 및 잠재적 오염 물질과 수술 부위의 오염 방지하기 위해 필요한 단계입니다. </li></ol> 2. 수술 <ol><li> 마취의 전체 기간 동안 따뜻함을위한 외부 열원을 제공합니다. 마취는 체온 조절의 장애를 야기하기 때문에 동물은 일부 저체온증이 발생할 수 있습니다; 따라서, 체온을 유지한다. </li><li> 수술을 통해 그 뒷면에 마우스를 놓습니다. </li><li> 외과 테이프 조각과 수술 테이블에 앞 사지, 뒷다리와 마우스의 꼬리를 수정합니다. </li><li> xyphisternum에 직선 절개 즉시 열등합니다. </li><li> 간을 노출 멸균 자기 유지 외과 견인기를 사용합니다. </li><li> 주사기 차분히, 20G 바늘을 갖는 5 × 10 6 BNL 1ME A.7R.1를 포함하는 PBS 100 ㎕를 주입간 우엽에 마우스 간암 세포. </li><li> 바늘을 제거한 후, 인해 간 높은 혈관에 잠재적 인 출혈을 멈추게하기 위해 적어도 1 분 동안 멸균 면봉을 적용합니다. </li><li> 수술 클립을 사용하여 피부 폐쇄 다음 흡수성 봉합사와 피하 조직을 닫습니다. 그 후, betadine 스크럽과 함께 다시 한 번 상처를 청소하십시오. 참고 :이 수술 상처의 폐쇄를 보장합니다. </li><li> 피하 생리 식염수 100 ㎕를 제공합니다. </li></ol> 3. 수술 후 <ol><li> 수술 후 즉시, 마우스 조건 관리인에게 통지 수술 후 신분증 깨끗한 케이지에 마우스를 옮긴다. 음식과 물을 제공합니다. 참고 : 우리의 경험에 의하면, 초기 24 시간의 기간 후 수술 젖은 음식과 물을 혼자 보관하면 그냥 최선을 운임됩니다 수술을 한 마우스를. </li><li> 38 ºC을 능가하지 열을 생성하는 가열 램프와 함께 동물을 따뜻하게 유지. 유마우스가 완전히 회복 될 때까지 온도를 측정하는 온도계를 SE는. </li><li> 바람직하게는 종이 침구와 케이지 하우스 동물의 편안함을 보장합니다. </li><li> 초기 복구 한 후, 불편 함, 출혈, 통증과 고통의 징후 동물을 모니터링 할 수 있습니다. 통증을 완화하기 위해 프레 노르 핀 (0.05 -0.1 ㎎ / ㎏)을 얻었다. 밀접하게 약 2 시간 동안 활성 동작과 모양뿐만 아니라 물과 음식 소비를 모니터링. 외래 경우, 마우스의 정규 일반 주택을 다시 지역에 동물을 이동하고 24 시간 후 수술 6-12 시간 간격으로 모니터링을 계속합니다. 원하는 경우 체중 검사를 수행합니다. </li><li> 동물이 문제가 회복 데 경우, 적절한지지 요법의 조치를 취할. 마우스는 복구 할 수없는 경우, 인도적으로 CO 2 질식 자궁 경부 전위를 통해 쥐를 안락사. 안락사에 대한 승인 기관의 지침을 따르십시오. </li><li> 수술 후 보통 observab입니다 간세포 암의 임상 적 증거를 확인몸 상태 점수 르로 크게 감소. 주 :이 시점에서, 일차 종양 가능성 간에서 개발 및 전이성 예금 가능성 폐에서 개발 하였다. </li><li> 이 시점에서, 인도적인 안락사에 의해 실험에 참여 (실험 및 제어) 모든 쥐를 희생. 이산화탄소로 질식 생쥐 대상 : 매우 천천히 -10 5 분에 걸쳐 챔버 내에 이산화탄소를 방출. 이것은 호흡 정지의 원인이됩니다. </li></ol>

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The authors declare that they have no competing financial interests.

이 문서의 방법에 대한 깊이있는 설명은 (그림 2)이 최근 Ogunwobi 및 간세포 암 전이의 동소 동계 마우스 모델의 성공적인 설립의 동료에 의해보고되었다 주어진다 3. 이 절차에 대한 종양의 테이크 속도는 일반적으로 높다. 우리는 이전에 100 % 3의 종양 걸릴 속도를 관찰했다. 그러나 테이크 속도는 연구자의 능력에 따라 변수가 될 수 있습니다. 훈련하에 새로운 대학원?...

Hepatocellular carcinoma (HCC) is one of the most lethal cancers, with poor prognosis and low life expectancy. Nearly all cancer-related deaths are due to the metastatic spread of the disease from the originating organ to additional distant organs2-6.HCC progression is a complex process. Therefore, being able to model the tumor microenvironment that is naturally found in metastatic HCC in animal models can prove to be a successful and useful way to reveal relevant mechanisms in humans.7Unfortunately, the mechanisms of cancer metastasis are still largely unclear. Therefore, there is a need to establish animal models that will enable us to elucidate the underlying molecular mechanisms of metastases of cancers such as HCC3,5.
Mouse model systems are a very useful approach for delineating mechanisms and evaluating novel strategies for treatment of metastatic human cancers5,8. The various mouse model systems that presently exist are a testament to efforts of researchers to correctly depict the complexity of the disease5,8,9.
Ogunwobi and colleagues recently used a survival surgical approach to demonstrate establishment of a novel orthotopic syngeneic mouse model for the study of metastasis in HCC3. Their work established a &#8220;patient-like&#8221; mouse model that recapitulates features of aggressive and metastatic HCCs3. They further demonstrated that this mouse model system can be used to study the biology of circulating tumor cells, and that this holds potential for gaining novel insights into the mechanisms of cancer metastasis3.
The aim of this paper is to describe in detail the methodology used in establishing this &#8220;patient-like&#8221; orthotopic syngeneic mouse model of HCC metastasis3. The methodology of how to implant BNL 1ME A.7R.1 mouse HCC cells directly into the liver (the organ of origin) of the inbred wild-type immune competent laboratory mouse strain, BALB/c3 using survival surgery will be described. Unlike other mouse xenograft tumor models where human tumor cells are implanted into immune deficient mice, this system is syngeneic and is, therefore, suitable for studying the role of the immune system in tumor metastasis3,5,8. This approach will likely gain widespread use for studying the mechanisms of metastasis in solid organ cancers.

<table><tbody><tr><td>Micro Dissecting Tweezer&amp;nbsp;</td><td>Roboz</td><td>(RS-5040)</td><td>Tip 0.20 x 0.12 mm</td></tr><tr><td>Graefe Micro Dissecting Forceps, serrated curved tip</td><td>Roboz</td><td>(RS-5111)</td><td>1 X 2 teeth, curved tip width 0.6 mm</td></tr><tr><td>Micro Dissecting Retractor-Agricola (3 by 3 prongs)</td><td>Roboz</td><td>(RS-6501)</td><td>Blunt 3 X 3 prongs, depth 4 mm, spread 25 mm</td></tr><tr><td>Micro Dissecting Retractor-Goldstein (3 by 3 prongs)</td><td>Roboz</td><td>(RS-6503)</td><td>Blunt, 3 X 3 prongs, depth 4 mm, spread 19 mm</td></tr><tr><td>Jameson Caliper (Measures tumors)</td><td>Roboz</td><td>(RS-6466)</td><td>80 mm/3 inch scale, chrome plated&amp;nbsp;</td></tr><tr><td>Micro Dissecting Scissors, large ring sissors, straight and sharp</td><td>Roboz</td><td>(RS-5852)</td><td>23 mm blades, length 4 inches, flat shanks and large rings</td></tr><tr><td>Scalpel with blades, for delicate dissecting procedures</td><td>Roboz</td><td>(RS-9861-36)</td><td></td></tr><tr><td>Scalpel handle&amp;nbsp;</td><td>Roboz</td><td>(RS-9884)</td><td>Solid</td></tr><tr><td>Littauer Stitch Sissors</td><td>Roboz</td><td>(RS-7074)</td><td>Length 4.5 inches</td></tr><tr><td>Brown needle holder, for easy suture tying</td><td>Roboz</td><td>(RS-7960)</td><td>Convex jaw, fine serrations</td></tr><tr><td>Reflex 7MM wound clips with reflex 7 clip applier</td><td>Roboz</td><td>(RS-9262)</td><td>safe, secure alternative method of wound closure</td></tr><tr><td>Instrument tray and lid</td><td>Roboz</td><td>RT-1350S</td><td></td></tr><tr><td>Mini-clipper with detachable blade</td><td>Roboz</td><td>(RS-5903)</td><td></td></tr><tr><td>Germinator 500 (the Germ Terminator) Dry Sterilizer</td><td>Roboz</td><td>Ds-400, Ds-401, DS-501</td><td>For fast decontamination of micro-dissecting instruments.&amp;nbsp;&amp;nbsp; Instruments decontaminate within 15 sec.&amp;nbsp;</td></tr><tr><td>Microinjection needles</td><td>VWR</td><td>BD305125</td><td>25G Needle</td></tr></tbody></table>

a "patient-like" orthotopic syngeneic mouse model of hepatocellular carcinoma metastasis

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of cancer metastasis are still unclear. Animal models are essential for elucidating the mechanisms and for evaluating novel strategies for the treatment of metastatic cancers. Here, an in-depth description of a &#8220;patient-like&#8221; orthotopic syngeneic mouse model for exploring the mechanisms of metastasis of solid organ tumors is provided. The survival surgical implantation of BNL 1ME A.7R.1 mouse hepatocellular carcinoma cells directly into the liver (the organ of origin) of the inbred wild-type immune competent laboratory mouse strain, BALB/c is described. The success and reproducibility of this methodology recommends it for widespread use in elucidating the biological mechanisms of solid organ cancer metastasis.

을 Balb / c 마우스의 간은 5 × 106 BNL 1ME A.7R.1 마우스 간암 세포를 이식 하였다. HCC 개발의 임상 적 증거가 육십삼일 수술 후 관찰했다. 따라서, 마우스는 인도적으로 안락사시켰다. 검시는 안락사 쥐에 대해 수행 하였다. 폐 및 간을 절제하고, 조심 육안 검사를 육안 종양을 확인하기 위해 수행 하였다. 하나의 마우스에서 종양이 표면 복벽 (도 1a)에 일부 첨부 간 표면에서 발견되...

Watch this Scientific Journal Video about A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis at JoVE.com

A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis

암의 전이 확산 암 관련 사망의 주요 원인이다. 우리는 고형 장기의 종양에서 전이의 메커니즘을 연구하기위한 &quot;환자와 같은&quot;동 소성 동계 마우스 모델 시스템 구축을위한 우리의 생존 수술 방법에 대한 심층적 인 설명을 제공한다.

&quot;환자와 같은&quot;동소 이식 동계 마우스 모델 간세포 암의 전이

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of ...

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12.2K 조회수.  Hunter College of the City University of New York.  암의 전이 확산 암 관련 사망의 주요 원인이다. 우리는 고형 장기의 종양에서 전이의 메커니즘을 연구하기위한 "환자와 같은"동 소성 동계 마우스 모델 시스템 구축을위한 우리의 생존 수술 방법에 대한 심층적 인 설명을 제공한다. 

비디오: A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis

Advanced Animal Model of Colorectal Metastasis in Liver: Imaging Techniques and Properties of Metastatic Clones

Patients with a limited number of hepatic metastases and slow rates of progression can be successfully treated with local treatment approaches1,2. However, little is known about the heterogeneity of liver metastases, and animal models capable of evaluating the development of individual metastatic colonies are needed. Here, we present an advanced model of hepatic metastases that provides the ability to quantitatively visualize the development of individual tumor clones in the liver and estimate their growth kinetics and colonization efficiency. We generated a panel of monoclonal derivatives of HCT116 human colorectal cancer cells stably labeled with luciferase and tdTomato and possessing different growth properties. With a splenic injection followed by a splenectomy, the majority of these clones are able to generate hepatic metastases, but with different frequencies of colonization and varying growth rates. Using the In Vivo&#160;Imaging System (IVIS), it is possible to visualize and quantify metastasis development with in vivo luminescent and ex vivo fluorescent imaging. In addition, Diffuse Luminescent Imaging Tomography (DLIT) provides a 3D distribution of liver metastases in vivo. Ex vivo fluorescent imaging of harvested livers provides quantitative measurements of individual hepatic metastatic colonies, allowing for the evaluation of the frequency of liver colonization and the growth kinetics of metastases. Since the model is similar to clinically observed liver metastases, it can serve as a modality for detecting genes associated with liver metastasis and for testing potential ablative or adjuvant treatments for liver metastatic disease.

The ability of metastatic clones to colonize distant sites depends on their proliferation capacity and/or their ability to survive in the host microenvironment without significant proliferation. Here, we present an animal model that allows quantitative visualization of both types of liver colonization by metastatic clones.

고급 동물의 간에서 대장 전이의 모델 : 이미징 기술 및 전이성 클론의 특성

Patients with a limited number of hepatic metastases and slow rates of progression can be successfully treated with local treatment approaches1,2. ...

연구

암 연구학

A Portal Vein Injection Model to Study Liver Metastasis of Breast Cancer

Breast cancer is the leading cause of cancer-related mortality in women worldwide. Liver metastasis is involved in upwards of 30% of cases with breast cancer metastasis, and results in poor outcomes with median survival rates of only 4.8 - 15 months. Current rodent models of breast cancer metastasis, including primary tumor cell xenograft and spontaneous tumor models, rarely metastasize to the liver. Intracardiac and intrasplenic injection models do result in liver metastases, however these models can be confounded by concomitant secondary-site metastasis, or by compromised immunity due to removal of the spleen to avoid tumor growth at the injection site. To address the need for improved liver metastasis models, a murine portal vein injection method that delivers tumor cells firstly and directly to the liver was developed. This model delivers tumor cells to the liver without complications of concurrent metastases in other organs or removal of the spleen. The optimized portal vein protocol employs small injection volumes of 5 - 10 &#956;l, &#8805; 32 gauge needles, and hemostatic gauze at the injection site to control for blood loss. The portal vein injection approach in Balb/c female mice using three syngeneic mammary tumor lines of varying metastatic potential was tested; high-metastatic 4T1 cells, moderate-metastatic D2A1 cells, and low-metastatic D2.OR cells. Concentrations of &#8804; 10,000 cells/injection results in a latency of ~ 20 - 40 days for development of liver metastases with the higher metastatic 4T1 and D2A1 lines, and &#62; 55 days for the less aggressive D2.OR line. This model represents an important tool to study breast cancer metastasis to the liver, and may be applicable to other cancers that frequently metastasize to the liver including colorectal and pancreatic adenocarcinomas.

A surgical procedure was developed to deliver mammary tumor cells to the murine liver via portal vein injection. This model permits investigation of late stages of liver metastasis in a fully immune competent host, including tumor cell extravasation, seeding, survival, and metastatic outgrowth in the liver.

포탈 정맥 주입 모델 간 전이 유방암의 공부를

Breast cancer is the leading cause of cancer-related mortality in women worldwide. Liver metastasis is involved in upwards of 30% of cases with breast ...

A Syngeneic Mouse Model of Metastatic Renal Cell Carcinoma for Quantitative and Longitudinal Assessment of Preclinical Therapies

Renal cell carcinoma (RCC) affects &#62; 60,000 people in the United States annually, and ~ 30% of RCC patients have multiple metastases at the time of diagnosis. Metastatic RCC (mRCC) is incurable, with a median survival time of only 18 months. Immune-based interventions (e.g., interferon (IFN) and interleukin (IL)-2) induce durable responses in a fraction of mRCC patients, and multikinase inhibitors (e.g., sunitinib or sorafenib) or anti-VEGF receptor monoclonal antibodies (mAb) are largely palliative, as complete remissions are rare. Such shortcomings in current therapies for mRCC patients provide the rationale for the development of novel treatment protocols. A key component in the preclinical testing of new therapies for mRCC is a suitable animal model. Beneficial features that recapitulate the human condition include a primary renal tumor, renal tumor metastases, and an intact immune system to investigate any therapy-driven immune effector responses and the formation of tumor-induced immunosuppressive factors. This report describes an orthotopic mRCC mouse model that has all of these features. We describe an intrarenal implantation technique using the mouse renal adenocarcinoma cell line Renca, followed by the assessment of tumor growth in the kidney (primary site) and lungs (metastatic site).

Implementation of an orthotopic model of renal cell carcinoma in immunocompetent mice affords the investigator a clinically-relevant system defined by the presence of a primary renal tumor and lung metastases in the same animal. This system can be used to preclinically test a variety of treatments in vivo.

전임상 치료의 양적 및 종 방향 평가를위한 동계 마우스 모델 전이성의 신장 세포 암

Renal cell carcinoma (RCC) affects &#62; 60,000 people in the United States annually, and ~ 30% of RCC patients have multiple metastases at the time of ...

Orthotopic Mouse Model of Colorectal Cancer

The traditional subcutaneous tumor model is less than ideal for studying colorectal cancer. Orthotopic mouse models of colorectal cancer, which feature cancer cells growing in their natural location, replicate human disease with high fidelity. Two techniques can be used to establish this model. Both techniques are similar and require mouse anesthesia and laparotomy for exposure of the cecum. One technique involves injection of a colorectal cancer cell suspension into the cecal wall. Cancer cells are first grown in culture, harvested when subconfluent and prepared as a single cell suspension. A small volume of cells is injected slowly to avoid leakage. The other technique involves transplantation of a piece of subcutaneous tumor onto the cecum. A mouse with a previously established subcutaneous colorectal tumor is euthanized and the tumor is removed using sterile technique. The tumor piece is divided into small pieces for transplantation to another mouse. Prior to transplantation, the cecal wall is lightly damaged to facilitate tumor cell infiltration. The time to developing primary tumors and liver metastases will vary depending on the technique, cell line, and mouse species used. This orthotopic mouse model is useful for studying the natural progression of colorectal cancer and testing new therapeutic agents against colorectal cancer.

Two techniques can be used to establish this model: injection of a cancer cell suspension into the cecal wall or transplantation of a piece of subcutaneous tumor onto the cecum. This model is useful for studying the natural progression of colorectal cancer and testing new therapeutic agents against colorectal cancer.

대장암의 Orthotopic 마우스 모델

The traditional subcutaneous tumor model is less than ideal for studying colorectal cancer. Orthotopic mouse models of colorectal cancer, which feature ...

생물학

An Oncogenic Hepatocyte-Induced Orthotopic Mouse Model of Hepatocellular Cancer Arising in the Setting of Hepatic Inflammation and Fibrosis

The absence of a clinically relevant animal model addressing the typical immune characteristics of hepatocellular cancer (HCC) has significantly impeded elucidation of the underlying mechanisms and development of innovative immunotherapeutic strategies. To develop an ideal animal model recapitulating human HCC, immunocompetent male C57BL/6J mice first receive a carbon tetrachloride (CCl4) injection to induce liver fibrosis, then receive histologically-normal oncogenic hepatocytes from young male SV40 T antigen (TAg)-transgenic mice (MTD2) by intra-splenic (ISPL) inoculation. Androgen generated in recipient male mice at puberty initiates TAg expression under control of a liver-specific promoter. As a result, the transferred hepatocytes become cancer cells and form tumor masses in the setting of liver fibrosis/cirrhosis. This novel model mimics human HCC initiation and progression in the context of liver fibrosis/cirrhosis and reflects the most typical features of human HCC including immune dysfunction.

Here, we describe the development of a clinically relevant murine model of liver cancer recapitulating the typical immune features of hepatocellular cancer (HCC).

간 염증과 섬유증의 설정에서 발생하는 간세포암의 종양 성 간세포 유도 직교 마우스 모델

The absence of a clinically relevant animal model addressing the typical immune characteristics of hepatocellular cancer (HCC) has significantly impeded ...

Murine Model of Metastatic Liver Tumors in the Setting of Ischemia Reperfusion Injury

Liver ischemia and reperfusion (I/R) injury, a common clinical challenge, remains an inevitable pathophysiological process that has been shown to induce multiple tissue and organ damage. Despite recent advances and therapeutic approaches, the overall morbidity has remained unsatisfactory especially in patients with underlying parenchymal abnormalities. In the context of aggressive cancer growth and metastasis, surgical I/R is suspected to be the promoter regulating tumor recurrence. This article aims to describe a clinically relevant murine model of liver I/R and colorectal liver metastasis. In doing so, we aim to assist other investigators in establishing and perfecting this model for their routine research practice to better understand the effects of liver I/R on promoting liver metastases.

We describe in detail a clinically relevant colorectal cancer liver metastases (CRLM) tumor model and the influence of liver ischemia reperfusion (I/R) in tumor growth and metastasis. This model can help to better understand the mechanisms underlying surgery-induced promotion of liver metastatic growth.

허혈 재관류 손상의 설정에서 전이성 간 종양의 뮤린 모델

Liver ischemia and reperfusion (I/R) injury, a common clinical challenge, remains an inevitable pathophysiological process that has been shown to induce ...

Generation of Subcutaneous and Intrahepatic Human Hepatocellular Carcinoma Xenografts in Immunodeficient Mice

In vivo experimental models of hepatocellular carcinoma (HCC) that recapitulate the human disease provide a valuable platform for research into disease pathophysiology and for the preclinical evaluation of novel therapies. We present a variety of methods to generate subcutaneous or orthotopic human HCC xenografts in immunodeficient mice that could be utilized in a variety of research applications. With a focus on the use of primary tumor tissue from patients undergoing surgical resection as a starting point, we describe the preparation of cell suspensions or tumor fragments for xenografting. We describe specific techniques to xenograft these tissues i) subcutaneously; or ii) intrahepatically, either by direct implantation of tumor cells or fragments into the liver, or indirectly by injection of cells into the mouse spleen. We also describe the use of partial resection of the native mouse liver at the time of xenografting as a strategy to induce a state of active liver regeneration in the recipient mouse that may facilitate the intrahepatic engraftment of primary human tumor cells. The expected results of these techniques are illustrated. The protocols described have been validated using primary human HCC samples and xenografts, which typically perform less robustly than the well-established human HCC cell lines that are widely used and frequently cited in the literature. In comparison with cell lines, we discuss factors which may contribute to the relatively low chance of primary HCC engraftment in xenotransplantation models and comment on technical issues that may influence the kinetics of xenograft growth. We also suggest methods that should be applied to ensure that xenografts obtained accurately resemble parent HCC tissues.

Human tumor xenografts in immunodeficient mice are valuable tools to study cancer biology. Specific protocols to generate subcutaneous and intrahepatic xenografts from human hepatocellular carcinoma cells or tumor fragments are described. Liver regeneration induced by partial hepatectomy in recipient mice is presented as a strategy to facilitate intrahepatic engraftment.

면역 결핍 마우스의 피하 및 간내 인간의 간세포 암 이종 이식의 생성

In  vivo experimental models of hepatocellular carcinoma (HCC) that recapitulate  the human disease provide a valuable platform for research into disease  ...

의학

Non-Invasive PET/MR Imaging in an Orthotopic Mouse Model of Hepatocellular Carcinoma

Preclinical experimental models of hepatocellular carcinoma (HCC) that recapitulate human disease represent an important tool to study tumorigenesis and evaluate novel therapeutic approaches. Non-invasive whole-body imaging using positron emission tomography (PET) provides critical insights into the in vivo characteristics of tissues at the molecular level in real-time. We present here a protocol for orthotopic HCC xenograft creation with and without hepatic artery ligation (HAL) to induce tumor hypoxia and the assessment of their tumor metabolism in vivo using [18F]Fluoromisonidazole ([18F]FMISO) and [18F]Fluorodeoxyglucose ([18F]FDG) PET/magnetic resonance (MR) imaging. Tumor hypoxia could be readily visualized using the hypoxia marker [18F]FMISO, and it was found that the [18F]FMISO uptake was higher in HCC mice that underwent HAL than in the non-HAL group, whereas [18F]FDG could not distinguish tumor hypoxia between the two groups. HAL tumors also displayed a higher level of hypoxia-inducible factor (HIF)-1&#945; expression in response to hypoxia. Quantification of HAL tumors showed a 2.3-fold increase in [18F]FMISO uptake based on the standardized value uptake (SUV) approach.

Here, we present a protocol to create orthotopic hepatocellular carcinoma xenografts with and without hepatic artery ligation and perform non-invasive positron emission tomography (PET) imaging of tumor hypoxia using [18F]Fluoromisonidazole ([18F]FMISO) and [18F]Fluorodeoxyglucose ([18F]FDG).

간세포 암종의 동소 마우스 모델에서 비침습적 PET/MR 이미징

Preclinical experimental models of hepatocellular carcinoma (HCC) that recapitulate human disease represent an important tool to study tumorigenesis and ...

A Preclinical Murine Model of Hepatic Metastases

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, murine pancreatic tumor model of hepatic metastases via a hemispleen injection of syngeneic murine pancreatic tumor cells is described. This model mimics many of the clinical conditions in patients with metastatic disease to the liver. Mice consistently develop metastases in the liver allowing for investigation of the metastatic process, experimental therapy testing, and tumor immunology research.

A preclinical, murine model of hepatic metastases performed via a hemispleen injection technique.

간 전이의 전임상 쥐과 모델

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

An Orthotopic Mouse Model of Spontaneous Breast Cancer Metastasis

Metastasis is the primary cause of mortality of breast cancer patients. The mechanism underlying cancer cell metastasis, including breast cancer metastasis, is largely unknown and is a focus in cancer research. Various breast cancer spontaneous metastasis mouse models have been established. Here, we report a simplified procedure to establish orthotopic transplanted breast cancer primary tumor and resultant spontaneous metastasis that mimic human breast cancer metastasis. Combined with the bioluminescence live tumor imaging, this mouse model allows tumor growth and progression kinetics to be monitored and quantified. In this model, a low dose (1 x 104 cells) of 4T1-Luc breast cancer cells was injected into BALB/c mouse mammary fat pad using a tuberculin syringe. Mice were injected with luciferin and imaged at various time points using a bioluminescent imaging system. When the primary tumors grew to the size limit as in the IACUC-approved protocol (approximately 30 days), mice were anesthetized under constant flow of 2% isoflurane and oxygen. The tumor area was sterilized with 70% ethanol. The mouse skin around the tumor was excised to expose the tumor which was removed with a pair of sterile scissors. Removal of the primary tumor extends the survival of the 4T-1 tumor-bearing mice for one month. The mice were then repeatedly imaged for metastatic tumor spreading to distant organs. Therapeutic agents can be administered to suppress tumor metastasis at this point. This model is simple and yet sensitive in quantifying breast cancer cell growth in the primary site and progression kinetics to distant organs, and thus is an excellent model for studying breast cancer growth and progression, and for testing anti-metastasis therapeutic and immunotherapeutic agents in vivo.

An orthotopic breast cancer primary tumor model and surgical removal of primary tumor to extend mouse life to generate spontaneous metastasis are described. The tumor growth and progression are monitored and quantified by luciferase fluorescence imaging.

"환자와 같은"동소 이식 동계 마우스 모델 간세포 암의 전이

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