Name: a preclinical murine model of hepatic metastases
Uploaded: 2014-09-27T13:00:00.000+00:00
Duration: 6 min 51 s
Description: Watch this Scientific Journal Video about A Preclinical Murine Model of Hepatic Metastases at JoVE.com

KF y KO son co primeros autores, junto con KS Este trabajo fue apoyado en parte por el AHPBA Fellowship (KS), NIH K23 CA148964-01 (LZ), la Escuela de Medicina Johns Hopkins premio Clinical Scientist (LZ), Fundación Viragh y el Centro de Cáncer Skip Viragh pancreático en la Universidad Johns Hopkins (EMJ y LZ), la Fundación Nacional de Páncreas (LZ), la Fundación Lefkofsky Familia (LZ), el NCI SPORE en Gastrointestinal Cánceres P50 CA062924 (EMJ, LZ), la Fundación Lustgarten (LZ), y las subvenciones del Centro de Cáncer Sol Goldman pancreático (KS , BHE, LZ).

Todos los experimentos con animales se ajustaban a las directrices del Comité para el Cuidado y Uso de Animales de la Universidad Johns Hopkins, y los animales fueron mantenidos de acuerdo con las directrices de la Asociación para la Evaluación y Acreditación de Cuidado de Laboratorio (AAALAC). El procedimiento quirúrgico se realiza bajo condiciones estériles en un quirófano que sufre un mínimo de quince cambios de aire por hora. Todos los instrumentos son esterilizados en autoclave antes del procedimiento y de nuevo con etanol al 70% entre cada ratón durante el procedimiento. Todos los ratones que no llegará a la intervención quirúrgica son sacrificados bajo CO 2 durante 3 minutos seguido por dislocación cervical, mientras que todavía en la anestesia. 1. Preparación de la célula <ol><li> Resuspender las células cultivadas en Panc02 fosfato salino tamponado a 2 x10 7 células / ml, y mantener en hielo. </li></ol> 2. Preparación Ratón <ol><li> Administrar ketaminae (100 mg / kg) mezclado con xilazina (10 mg / kg) por vía intraperitoneal a 8-12 semanas de edad C57BL / 6 ratones hembra en dosis divididas. </li><li> Compruebe que la punta retirada pizca reflejo se abolió para asegurarse de que el ratón esté totalmente anestesiado. </li><li> Administrar dosis adicionales de ketamina (100 mg / kg) mezclado con xilazina (10 mg / kg) según sea necesario para anestesiar totalmente los ratones. </li><li> Aplicar Puralube Vet Ungüento para los ojos de los ratones para que no se seque mientras que los ratones están bajo anestesia. </li><li> Afeitar el área quirúrgica de los ratones para evitar la contaminación de la herida. </li><li> Preparar el área subcostal izquierdo de la incisión con etanol al 70% y luego yodo. </li><li> Coloque un paño quirúrgico alrededor del abdomen para mantener la esterilidad. </li></ol> 3. laparotomía <ol><li> Hacer una incisión subcostal izquierda en línea con el oído izquierdo a través de la piel y a través del peritoneo usando un escalpelo. </li><li> Expresar el bazo a través de la incisión aplicando p digitales simultánearesión a lo largo de los aspectos craneal y caudal de la incisión. </li><li> Localizar los vasos sanguíneos esplénicos en el extremo inferior del bazo. </li><li> Divida el bazo mediante la colocación de dos medianas clips de ligadura Horizon en el centro del bazo teniendo cuidado de no dañar los vasos sanguíneos del bazo en los polos del bazo. </li><li> Coloque el polo superior del bazo de nuevo en el peritoneo para evitar la contaminación. </li></ol> Inyección 4. Tumor <ol><li> Elaborar 150 l de solución salina tamponada con fosfato en un 26 T x 5/8 &quot;de la jeringa. </li><li> Elaborar 100 l de Panc02 (2 x10 6 células) en la misma jeringa manteniendo la jeringa en posición vertical en todo momento durante la inyección. </li><li> Sumergir la aguja en una solución de etanol al 70%. La aguja se sumerge en etanol al 70% para asegurar la esterilidad. </li><li> Inyectar las células lentamente en el hemispleen expuesta asegurándose la jeringa se mantiene en posición vertical en todo momento. El bisel de la Syringe debe observarse en todo momento a través de la cápsula esplénica para evitar la inyección de las células en el bazo. Un blanqueamiento del bazo y los vasos sanguíneos se debe observar después de la inyección. </li><li> Elevar el bazo y aplicar un medio Horizonte clip debajo del bazo para ocluir los vasos sanguíneos esplénicos. </li><li> Aplicar un pequeño clip Horizonte al aspecto más distal del páncreas y vasos esplénicos. </li><li> Ligar el páncreas y los vasos esplénicos de la hemispleen cortando por encima de los clips Horizon. </li><li> Coloca el ratón en su &#39;lado, y el riego de la incisión con agua destilada. </li></ol> 5. abdominal Clausura y recuperación de la anestesia <ol><li> Cierre del peritoneo con una puntada 4-0 consecutivo. </li><li> Aplicar una a dos clips de la piel para cerrar la incisión de la piel. </li><li> Administrar 0,1 mg / kg de buprenorfina o 5-10 mg / kg por vía subcutánea Carprofeno para aliviar el dolor post quirúrgico. </li><li> Coloca el ratón sobre una almohadilla térmica para recovery hasta móvil y el ratón demuestra patrones de respiración regulares. Los animales sólo se devuelven a la compañía de otros animales después de recuperarse plenamente de la intervención quirúrgica. </li></ol> 6. Necropsia Examen y cosecha del Hígado <ol><li> Entre 30 y 60 días después de la cirugía, los ratones comienzan a mostrar síntomas clínicos de metástasis y requerirán la eutanasia. Los signos de la enfermedad metastásica que requieren eutanasia humanitaria incluyen abdómenes agrandados y el desarrollo de ascitis. La eutanasia a los ratones por inhalación de CO 2. </li><li> Después de la inhalación, realice dislocación cervical. </li><li> Después de que el animal ha sido determinado que no responde, hacer una incisión a través del peritoneo para exponer el abdomen usando tijeras. </li><li> El uso de pinzas y tijeras, corte suavemente el hígado fuera del abdomen. </li><li> Coloque el hígado en compuesto OCT, y flash congelar con nitrógeno líquido. Guarde el hígado a -80 ° C. Alternativamente, el vivor puede fijarse en 16% neutros formalina tamponada a temperatura ambiente durante 24 horas y embebidos en parafina. </li></ol>

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Los autores no tienen conflicto de interés relevante para revelar.

Este modelo murino preclínica de la metástasis del cáncer de páncreas al hígado a través de una técnica de inyección hemispleen es el primer modelo descrito que refleja muchas de las condiciones clínicas e inmunológicas de los pacientes con enfermedad metastásica en el hígado. Este modelo ofrece varias ventajas sobre otros modelos murinos. En primer lugar, a diferencia de modelos transgénicos de cáncer de páncreas en la que sólo el 30% de los ratones desarrollan metástasis en el hígado y el momento de ...

Adenocarcinoma ductal pancreático (PDA) es la cuarta causa principal de muertes relacionadas con el cáncer en los Estados Unidos 1. La supervivencia de cinco años para los pacientes con PDA metastásico es 2-12% 1. Aunque la quimioterapia adyuvante y neoadyuvante para el cáncer de páncreas es hoy más eficaz que nunca, sigue existiendo una necesidad desesperada de nuevas terapias. El desarrollo de nuevas terapias requiere modelos animales preclínicos fiables. Aunque los modelos de tumores subcutáneos son sencillos de usar, inconvenientes incluyen la incapacidad de estos tumores y metástasis a imitar la progresión tumoral y microambientes visto en los cánceres humanos. Modelos de ingeniería genética, tales como el ratón ingeniería genética que contiene Kras y p53 knock-in mutaciones oncogénicas (el modelo KPC) 2 y el modelo KPC con un trazador de linaje YFP (el modelo PKCY 3) permiten el estudio de tumores que ocurren naturalmente en immunocompeteratones NT. Además, el microambiente tumoral se altera y se asemeja más estrechamente a la observada en la progresión tumoral humano. Por desgracia, el calendario de desarrollo de tumores es variable dentro de estos modelos, lo que hace difícil el estudio de terapias experimentales. Además, el retrocruzamiento de estos ratones requiere una cantidad significativa de tiempo y compromiso financiero, que no siempre es factible. Por último, los modelos de PDA xenoinjerto de ratón implantadas, requieran ratones, que han alterado o microambientes tumorales ausentes inmunocomprometidos. Como resultado, la eficacia de terapias experimentales demostrado en estos modelos preclínicos a menudo no son reproducibles en ensayos clínicos humanos 4. Este modelo de ratón hemisplenectomy utiliza células tumorales Panc02, que son una, inducida por metilcolantreno de páncreas línea celular de tumor altamente tumorigénico derivado de ratones C57BL / 6 5, 6. Además, otras líneas celulares PDA murinos derivan del micrófono KPCe se puede utilizar en este modelo. Schulick et al. Han desarrollado previamente una técnica hemisplenectomy y ha creado un modelo de ratón singénico de metástasis de cáncer de colon 7. Este modelo hemisplenectomy (Figura 1) ofrece la inoculación del tumor consistente e iguales en ratones inmunocompetentes se presta a la investigación de nuevos agentes terapéuticos 7-10.

<table><tbody><tr><td>Cell culture media and components</td><td></td><td></td><td>will vary depending on cell line</td></tr><tr><td>Phosphate Buffered Saline</td><td>Gibco by Life Technologies</td><td>10010-023</td><td></td></tr><tr><td>15 ml centrifuge tubes</td><td>Corning</td><td>430052</td><td></td></tr><tr><td>Curved Operating Scissor</td><td>Roboz Surgical Store</td><td>RS-6835</td><td></td></tr><tr><td>Micro Dissecting Graefe Forceps</td><td>Roboz Surgical Store</td><td>RS-5130</td><td></td></tr><tr><td>Needle holder (regular)</td><td>World Precision Instruments, Inc</td><td></td><td></td></tr><tr><td>3-0 or 4-0 suture</td><td></td><td></td><td>courtesy of dept of surgery, Johns Hopkins</td></tr><tr><td>Weck Horizon Open Ligating Clip Applier, small, angled</td><td>Teleflex</td><td>137085</td><td></td></tr><tr><td>Weck Horizon Open Ligating Clip Applier, medium, angled</td><td>Teleflex</td><td>237115</td><td></td></tr><tr><td>Weck Horizon medium ligating clips</td><td>Teleflex</td><td>002200</td><td></td></tr><tr><td>Weck Horizon small ligating clips</td><td>Teleflex</td><td>001200</td><td></td></tr><tr><td>Syringe, 1 cc, tb syringe, 3/8&quot; slip tip</td><td>Becton Dickinson</td><td>309625</td><td></td></tr><tr><td>Syringe, 1 cc, tb syringe, 5/8&quot; slip tip</td><td>Becton Dickinson</td><td>309597</td><td></td></tr><tr><td>9 mm Autoclip Applier</td><td>Mikron</td><td>427630</td><td></td></tr><tr><td>9 mm Autoclip</td><td>Becton Dickinson</td><td>427631</td><td></td></tr><tr><td>Heating pad</td><td>Sunbeam</td><td>CAT93</td><td></td></tr><tr><td>70% Ethanol</td><td></td><td></td><td>sold by numerous vendors</td></tr><tr><td>Ketamine Hydrochloride&amp;nbsp;</td><td>Hospira</td><td>22395DD</td><td></td></tr><tr><td>Xylazine hydrochloride</td><td>Sigma</td><td>X1251</td><td></td></tr></tbody></table>

a preclinical murine model of hepatic metastases

Se han desarrollado numerosos modelos murinos para estudiar los cánceres humanos y avanzar en la comprensión de tratamiento y desarrollo del cáncer. Aquí, se describe un modelo preclínico, de páncreas murino tumoral de las metástasis hepáticas por medio de una inyección hemispleen de las células tumorales pancreáticas murinos singénicos. Este modelo imita muchas de las condiciones clínicas en pacientes con enfermedad metastásica en el hígado. Los ratones desarrollan consistentemente metástasis en el hígado permitiendo para la investigación del proceso metastásico, las pruebas de terapia experimental, la investigación y la inmunología tumoral.

Panc02 células tumorales inyectadas en el hemispleen (Figura 1) metástasis de hígado en forma 100% de los ratones, mientras que de pulmón y las metástasis peritoneales no se observan si la técnica se realiza correctamente. Esta técnica se ha realizado en más de un centenar de ratones usando células Panc02 en múltiples experimentos independientes, y reproducible, todos los ratones no tratados mueren con metástasis hepáticas de entre 30 y 60 días después del procedimiento hemisplenectomy <st...

Watch this Scientific Journal Video about A Preclinical Murine Model of Hepatic Metastases at JoVE.com

A Preclinical Murine Model of Hepatic Metastases

Un modelo preclínico, murino de metástasis hepáticas realiza a través de una técnica de inyección hemispleen.

A preclínicos modelo murino de metástasis hepática

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

a-preclinical-murine-model-of-hepatic-metastases

Research

JoVE Journal

Medicine

28.6K vistas.  The Johns Hopkins University School of Medicine.  Un modelo preclínico, murino de metástasis hepáticas realiza a través de una técnica de inyección hemispleen. 

Video: A Preclinical Murine Model of Hepatic Metastases

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.

Murino hepática Bioluminiscente modelo tumoral

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

Investigación

Medicina

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.

Avanzada modelo animal de colorrectal metástasis en hígado: Técnicas de imagen y propiedades metastásicas de los clones

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

Investigación sobre el cáncer

Surgical Procedures and Methodology for a Preclinical Murine Model of De Novo Mammary Cancer Metastasis

A rate-limiting aspect of transgenic mouse models of mammary adenocarcinoma is that primary tumor burden in mammary tissue typically defines study end-points. Thus, studies focused on elucidating mechanisms of late-stage de novo metastasis are compromised, as are studies examining efficacy of anti-cancer therapies targeting mediators of metastasis in the adjuvant setting. Numerous murine mammary cancer models have been developed via targeted expression of dominant oncoproteins to mammary epithelial cells yielding models variably mimicking histopathologic and transcriptome-defined breast cancer subtypes common in women1. While much has been learned regarding the biology of mammary carcinogenesis with these models, their utility in identifying molecules regulating growth of late-stage metastasis are compromised as mice are typically euthanized at earlier time points due to significant primary tumor burden. Moreover, since a significant percentage of women diagnosed with breast cancer receive adjuvant therapy after surgical resection of primary tumors and prior to presence of detectable metastatic disease, preclinical models of de novo metastasis are urgently needed as platforms to evaluate new therapies aimed at targeting metastatic foci. To address these deficiencies, we developed a murine model of de novo mammary cancer metastasis, wherein primary mammary tumors are surgically resected, and metastatic foci subsequently develop over a 115 day post-surgical period. This long latency provides a tractable model to identify functionally significant regulators of metastatic progression in mice lacking primary tumor, as well as a model to evaluate preclinical therapeutic efficacy of agents aimed at blocking functionally significant molecules aiding metastatic tumor survival and growth.

Surgical Procedures and Methodology for a Preclinical Murine Model of De Novo Mammary Cancer Metastasis

Pre-clinical models evaluating adjuvant therapy targeting breast cancer metastasis are lacking. To address this, we developed a murine model of de novo pulmonary mammary adenocarcinoma metastasis, wherein therapies administered in the adjuvant setting (post surgical resection of primary tumors) can be evaluated for efficacy in impacting previously seeded pulmonary metastases.

Procedimientos quirúrgicos y metodología para un modelo murino preclínico<em&gt; De Novo</em&gt; Metástasis del cáncer mamario

A rate-limiting aspect of transgenic mouse models of mammary adenocarcinoma is that primary tumor burden in mammary tissue typically defines study ...

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.

Una inyección Modelo vena porta del hígado para estudiar la metástasis del cáncer de mama

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 ...

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).

Un modelo de ratón ortotópico inducido por hepatocitos oncogénicos que surge en el ajuste de la inflamación hepática y la fibrosis

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.

Modelo murino de tumores hepáticos metastásicos en el escenario de la lesión por reperfusión de isquemia

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

Portal Vein Injection of Colorectal Cancer Organoids to Study the Liver Metastasis Stroma

Hepatic metastasis of colorectal cancer (CRC) is a leading cause of cancer-related death. Cancer-associated fibroblasts (CAFs), a major component of the tumor microenvironment, play a crucial role in metastatic CRC progression and predict poor patient prognosis. However, there is a lack of satisfactory mouse models to study the crosstalk between metastatic cancer cells and CAFs. Here, we present a method to investigate how liver metastasis progression is regulated by the metastatic niche and possibly could be restrained by stroma-directed therapy. Portal vein injection of CRC organoids generated a desmoplastic reaction, which faithfully recapitulated the fibroblast-rich histology of human CRC liver metastases. This model was tissue-specific with a higher tumor burden in the liver when compared to an intra-splenic injection model, simplifying mouse survival analyses. By injecting luciferase-expressing tumor organoids, tumor growth kinetics could be monitored by in vivo imaging. Moreover, this preclinical model provides a useful platform to assess the efficacy of therapeutics targeting the tumor mesenchyme. We describe methods to examine whether adeno-associated virus-mediated delivery of a tumor-inhibiting stromal gene to hepatocytes could remodel the tumor microenvironment and improve mouse survival. This approach enables the development and assessment of novel therapeutic strategies to inhibit hepatic metastasis of CRC.

Portal vein injection of colorectal cancer (CRC) organoids generates stroma-rich liver metastasis. This mouse model of CRC hepatic metastasis represents a useful tool to study tumor-stroma interactions and develop novel stroma-directed therapeutics such as adeno-associated virus-mediated gene therapies.

Inyección en la vena porta de organoides de cáncer colorrectal para estudiar la metástasis hepática del estroma

Hepatic metastasis of colorectal cancer (CRC) is a leading cause of cancer-related death. Cancer-associated fibroblasts (CAFs), a major component of the ...

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.

Un modelo murino de cáncer de próstata humano Metástasis ortotópico

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  ...

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).

Imágenes PET/MR no invasivas en un modelo ortotópico de ratón de carcinoma hepatocelular

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

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.

The metastatic spread of cancer is the major cause of cancer-related deaths. We provide an in-depth description of our survival surgery methodology for establishing a &#8220;patient-like&#8221; orthotopic syngeneic mouse model system for studying the mechanisms of metastasis in solid organ tumors.

Un &quot;paciente-Like&quot; ortotópico Singénico modelo de ratón de carcinoma hepatocelular Metástasis

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 ...

A preclínicos modelo murino de metástasis hepática

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