Este trabajo fue financiado por el premio Nacional de Ciencias CARRERA Foundation (CBET-0846395) y el Departamento de Medicina Comparada, Universidad de Tulane.

1. Los animales <ol><li> Ratones hembra C57BL/6J (Mus musculus) se verifican libres de ectoparásitos contagiosa, endoparásitos helmintos, y los anticuerpos a 17 virus murinos por el proveedor antes del envío. Los ratones se encuentran en las instalaciones de animales AAALAC acreditado por la Universidad de Tulane, de acuerdo con la Guía para el Cuidado y Uso de Animales de Laboratorio 1. </li><li> En concreto, los ratones son alojados en grupo (4-5 ratones / jaula) en forma individual con ventilación tapas de policarbonato microaislamiento cageswith filtro de cama de madera dura. Se les proporciona alimentación ad libitum de roedores granulado y el agua del grifo. </li><li> Los ratones se les permite de 7 días para aclimatarse antes del inicio del estudio. </li></ol> 2. Las células <ol><li> El carcinoma de células transicionales murino línea celular MB49 se utiliza para producir los tumores de vejiga. </li><li> Las células se cultivaron en medio Dulbecco modificado de Eagle suplementado con suero fetal bovino al 10%, 100 unidades / ml de penicilina, y 100 unidades / ml de estreptomicina. </li><li> Cultivos se mantienen a 37 º C y 5% de CO 2 con los medios de comunicación cambia cada dos o tres días. </li></ol> 3. Animales modelo de tumor ortotópico La instilación de la modelo de tumor de vejiga se describe aquí es una modificación del procedimiento descrito por Gunther et al 2. <ol><li> La anestesia se indujo con 2% de isoflurano (MWI / VetOne, Meridian, ID, Cat. # 501017) en el oxígeno y se mantiene en 0,75-1,25%. (Varios animales pueden ser tratados al mismo tiempo, lo que exigiría un aumento en los niveles de isoflurano.) Para evitar que se seque la córnea, los ojos están lubricados con un ungüento oftálmico lágrimas artificiales. El calor del cuerpo se mantiene con un cojín calentado bajo una cortina de papel. </li><li> La piel de la espalda baja se recorta para promover el contacto con la placa de puesta a tierra de metal, y la zona de afeitado está recubierta con gel electroconduction. </li><li> Cada ratón se colocan individualmente en recumbancy dorsal sobre una placa de metal a tierra, y la vejiga se sondaje transuretral con un lubricado de calibre 24, 3 / 4 pulgadas, el exceso de la aguja del catéter IV con el estilete eliminado. </li><li> La vejiga urinaria se vacía de orina, y un alambre de platino se inserta a través del catéter en la vejiga urinaria dejando el cable expuesto en cada extremo de la sonda - de aproximadamente 2 mm en el extremo de la luz del catéter. El cable es de platino debido a la resistencia del material a la oxidación. </li><li> Una corriente de 2,5 W se aplica luego a la vejiga durante aproximadamente 0,5 segundos al tocar la parte exterior del cable con una unidad de electrocauterio establecido en la coagulación. </li><li> El alambre se mueve entonces a un segundo lugar en la vejiga y otra carga de 2,5 W aplicada. </li><li> Se retira el alambre, y 100 l de MB49 células a una concentración de 1x10 6 células / ml se inculca a través del catéter en la vejiga a través de una jeringa de 1 ml. </li><li> Las jeringas son colocada en los catéteres para evitar flujo de salida de las suspensiones celulares, y los ratones sondaje se mantienen bajo anestesia durante 90 minutos para permitir que las células se unen a los sitios de quema. </li><li> 100-200 l de solución salina normal se administra por inyección IP a cada ratón antes de la recuperación. </li><li> Al final del período de incubación de los catéteres se retiran y los animales se les permitió recuperarse en el teclado se calienta hasta reflejos de enderezamiento vuelta, momento en el que los ratones son devueltos a sus jaulas. </li></ol> 4. Ultrasonido <ol><li> Los tumores se dejaron crecer durante 3-5 días antes de los ratones someterse a un examen de ultrasonido de la vejiga urinaria para monitorizar el crecimiento del tumor. </li><li> Para los exámenes de ultrasonido inicial, los animales son anestesiados como se describió anteriormente. Los ratones se colocan en recumbancy dorsal, y la piel del abdomen se prepara recorte de la piel. </li><li> Un catéter de calibre 24 IV se inserta en transuretral de la vejiga urinaria. En este punto, la vejiga puede ser drenado y la orina recogida para su posterior análisis. </li><li> 100 l de solución salina estéril se instila en la vejiga a través del catéter para mejorar la visibilidad de la vejiga en la ecografía. </li><li> Gel para ultrasonido de conducción se aplica en el abdomen y la ecografía se realiza para evaluar la pared de la vejiga y de la luz. </li></ol> 5. Los resultados representativos: Tejido del tumor comienza a aparecer 3-5 días después de la inoculación. En nuestra experiencia, en un grupo de 12 ratones, 50% o más habitualmente tienen una masa de tejido detectable dentro de la vejiga (0,5 hasta 1,0 l, con V = π / 6 · L · W · h) por cinco días. Cuando se utiliza MB 49 células, los tumores seguirá creciendo y la eutanasia debe ser enviado hasta el día 24 si los ratones sin tratar. La eutanasia se recomienda cuando los ratones se letárgico o caquexia, la pantalla de pieles erizadas, encorvado, y el comportamiento no responde, o cuando pierden el 20% de su peso corporal inicial. &lt;/ P&gt; Que los tumores crecen rápidamente y tienen centros de necrosis después de llegar a una masa de 200 mg o más, 3 ostensiblemente porque las células en el tumor en desarrollo proliferan más rápido de lo que puede ser reparado por el suministro de sangre angiogénico. También se puede controlar indirectamente la progresión del tumor a través del nivel de hematurea, que está presumiblemente relacionada con la angiogénesis en los tumores de rápido desarrollo. Se observó consistentemente que la orina de tratar ratones con tumores restos de sangre el día 14. La concentración de los glóbulos rojos parece aumentar con el tiempo en estos ratones, y los volúmenes de orina recogida disminuido. Los tumores de más de 50 mg también se puede palpar con relativa facilidad, por lo general por 14 días después de la inoculación.

<ol>
	<li>Institute of Laboratory Animal Resources. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Guide for the Care and Use of Laboratory Animals. , (1996).</li><li>Gunther, J. H., Jurczok, A., Wulf, T., Brandau, S., Deinert, I., Jocham, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimizing+syngeneic+orthotopic+murine+bladder+cancer+(MB49).">Optimizing syngeneic orthotopic murine bladder cancer (MB49).</a> Cancer Res. 59, 2834-2837 (1999).</li><li>Zhang, X., Atala, A., Godbey, W. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression-targeted+gene+therapy+for+the+treatment+of+transitional+cell+carcinoma.">Expression-targeted gene therapy for the treatment of transitional cell carcinoma.</a> Cancer Gene Ther. 15, 543-552 (2008).</li></ol>

Los experimentos con animales se realizaron de acuerdo con las directrices y normas establecidas por el cuidado institucional y el empleo de la Universidad de Tulane.

Este método mínimamente invasivo para el establecimiento de los tumores de vejiga ortotópica en ratones hembras también se puede aplicar a otros tipos de células de MB49, a pesar de los diferentes resultados se deben anticipar. Al menos agresiva células tumorales se utilizan, aumentando ligeramente la potencia (con incrementos de 0,5 W para establecer el nuevo óptimo) rendirá un área más susceptible a la conexión de las células tumorales, y los tiempos de incubación de la vejiga para que las células inculc...

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<td>C57Bl/6J mice</td>
<td>&nbsp;</td>
<td>Jackson Laboratories</td>
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<td>Polycarbonate cages and ventilated rack</td>
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<td>Super Mouse 750</td>
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<td>Harlan Teklad</td>
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an orthotopic model of murine bladder cancer

En este sencillo procedimiento, los tumores de vejiga se establecen en las mujeres ratones C57 a través del uso de la cateterización, la cauterización local, y la adhesión celular posterior. Después de su vejiga está transuretral sondaje y drenaje, los animales son una vez más por sondaje para permitir la inserción de un alambre de platino en la vejiga sin dañar la uretra o la vejiga. Los catéteres están hechos de teflón para servir como aislante para los cables, que conducen la corriente eléctrica en la vejiga para crear una lesión por quemadura. Una unidad de electrocauterio son utilizados para entregar 2,5 W para el extremo expuesto del cable, quemando capas extracelular y proporcionar sitios de unión de las células de carcinoma que se entregan en la suspensión de la vejiga a través de un cateterismo posterior. Las células permanecen en la vejiga durante 90 minutos, tras lo cual los catéteres se retiran y la vejiga se deja escurrir de manera natural. El desarrollo de tumor se controla a través de ultrasonido. Se presta especial atención a la técnica de cateterismo en el video que lo acompaña.

Watch this Scientific Journal Video about An Orthotopic Model of Murine Bladder Cancer at JoVE.com

An Orthotopic Model of Murine Bladder Cancer

Los tumores de vejiga se establecen en las hembras de una forma mínimamente invasiva a través de cateterismo, la cauterización local, y posterior adhesión de las células cancerosas a los sitios de quema.

Un modelo ortotópico de cáncer de vejiga murino

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

an-orthotopic-model-of-murine-bladder-cancer

Research

JoVE Journal

Medicine

15.8K vistas.  Tulane University. Los tumores de vejiga se establecen en las hembras de una forma mínimamente invasiva a través de cateterismo, la cauterización local, y posterior adhesión de las células cancerosas a los sitios de quema.

Video: An Orthotopic Model of Murine Bladder Cancer

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.

Multi-fotón imágenes de invasión de células tumorales en un modelo de ratón ortotópico de carcinoma oral de células escamosas

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

Investigación

Medicina

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.

Combinación de radioterapia en un modelo ortotópico de tumor cerebral del ratón

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

Development of Obliterative Bronchiolitis in a Murine Model of Orthotopic Lung Transplantation

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and standardized not only for the study of allo-rejection but also between syngeneic strains for examining mechanisms of ischemia-reperfusion injury after lung transplantation. Although the application of the rat and other large animal model 2 contributed significantly to the elucidation of these studies, the scope of those investigations is limited by the scarcity of knockout and transgenic rats. Due to no effective therapies for obliterative bronchiolitis, the leading cause of death in lung transplant patients, there has been an intensive search for pre-clinical models that replicate obliterative bronchiolitis. The tracheal allograft model is the most widely used and may reproduce some of the histopathologic features of obliterative bronchiolitis 3. However, the lack of an intact vasculature with no connection to the recipient's conducting airways, and incomplete pathologic features of obliterative bronchiolitis limit the utility of this model 4. Unlike transplantation of other solid organs, vascularized mouse lung transplants have only recently been reported by Okazaki and colleagues for the first time in 2007 5. Applying the basic principles of the rat lung transplant, our lab initiated the obliterative bronchiolitis model using minor histoincompatible antigen murine orthotopic single-left lung transplants which allows the further study of obliterative bronchiolitis immunopathogenesis6.

Obliterative bronchiolitis is the key impediment to the long-term survival of lung transplant recipients and the lack of a robust preclinical model precludes examining obliterative bronchiolitis immunopathogenesis. Unlike other solid organ transplants, vascularized mouse lung transplantation has only recently been developed. Here we show our independently developed obliterative bronchiolitis model after murine orthotopic single-lung transplantation.

El desarrollo de bronquiolitis obliterante en un modelo murino de trasplante de pulmón ortotópico

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and ...

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.

Un modelo de tumor vesical ortotópica y la Evaluación de tratamiento intravesical SARNA

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

An Orthotopic Mouse Model of Anaplastic Thyroid Carcinoma

Several types of animal models of human thyroid carcinomas have been established, including subcutaneous xenograft and orthotopic implantation of cancer cells into immunodeficient mice. Subcutaneous xenograft models have been valuable for preclinical screening and evaluation of new therapeutic treatments. There are a number of advantages to using a subcutaneous model; 1) rapid, 2) reproducible, and 3) tumor establishment, growth, and response to therapeutic agents may be monitored by visual inspection. However, substantial evidence has shed light on the short-comings of subcutaneous xenograft models1-3. For instance, medicinal treatments demonstrating curative properties in subcutaneous xenograft models often have no notable impact on the human disease. The microenvironment of the site of xenographic transplantation or injection lies at the heart of this dissimilarity.
Orthotopic tumor xenograft models provide a more biologically relevant context in which to study the disease. The advantages of implanting diseased cells or tissue into their anatomical origin equivalent within a host animal includes a suitable site for tumor-host interactions, development of disease-related metastases and the ability to examine site-specific influence on investigational therapeutic remedies. Therefore, orthotopic xenograft models harbor far more clinical value because they closely reproduce human disease. For these reasons, a number of groups have taken advantage of an orthotopic thyroid cancer model as a research tool4-7.
Here, we describe an approach that establishes an orthotopic model for the study of anaplastic thyroid carcinoma (ATC), which is highly invasive, resists treatment, and is virtually fatal in all diagnosed patients. Cultured ATC cells are prepared as a dissociated cellular suspension in a solution containing a basement membrane matrix. A small volume is slowly injected into the right thyroid gland. Overall appearance and health of the mice are monitored to ensure minimal post-operative complications and to gauge pathological penetrance of the cancer. Mice are sacrificed at 4 weeks, and tissue is collected for histological analysis. Animals may be taken at later time-points to examine more advance progression of the disease. Production of this orthotopic mouse model establishes a platform that accomplishes two objectives: 1) further our understanding of ATC pathology, and 2) screen current and future therapeutic agents for efficacy in combating ATC.

Generation of an orthotopic mouse model of anaplastic thyroid carcinoma is described here. This technique employs surgical placement of human anaplastic thyroid cancer cells into the thyroid of immunodeficient mice, thus creating a more clinically relevant setting to study disease progression as well as screen innovative therapeutic interventions.

Un modelo de ratón ortotópico de carcinoma de tiroides anaplásico

Several types of animal models of human thyroid carcinomas have been established, including subcutaneous xenograft and orthotopic implantation of cancer ...

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.

Un modelo ortotópico de cáncer de vejiga de Estudios de liberación de genes

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

Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules that regulate HSCs. In these transplant model systems, the function of HSCs is determined by the ability of these cells to engraft and reconstitute lethally irradiated recipient mice. Commonly, the donor cell contribution/engraftment is measured by antibodies to donor- specific cell surface proteins using flow cytometry. However, this method heavily depends on the specificity and the ability of the cell surface marker to differentiate donor-derived cells from recipient-originated cells, which may not be available for all mouse strains. Considering the various backgrounds of genetically modified mouse strains in the market, this cell surface/ flow cytometry-based method has significant limitations especially in mouse strains that lack well-defined surface markers to separate donor cells from congenic recipient cells. Here, we reported a PCR-based technique to determine donor cell engraftment/contribution in transplant recipient mice. We transplanted male donor bone marrow HSCs to lethally irradiated congenic female mice. Peripheral blood samples were collected at different time points post transplantation. Bone marrow samples were obtained at the end of the experiments. Genomic DNA was isolated and the Y chromosome specific gene, Zfy1, was amplified using quantitative Real time PCR. The engraftment of male donor-derived cells in the female recipient mice was calculated against standard curve with known percentage of male vs. female DNAs. Bcl2 was used as a reference gene to normalize the total DNA amount. Our data suggested that this approach reliably determines donor cell engraftment and provides a useful, yet simple method in measuring hematopoietic cell reconstitution in murine bone marrow transplantation models. Our method can be routinely performed in most laboratories because no costly equipment such as flow cytometry is required.

Determining donor cell engraftment presents a challenge in mouse bone marrow transplant models that lack well-defined phenotypical markers. We described a methodology to quantify male donor cell engraftment in female transplant recipient mice. This method can be used in all mouse strains for the study of HSC functions.

Usando cuantitativa en tiempo real PCR para determinar quimerismo de células donantes en un modelo competitivo Trasplante de médula ósea murina

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules ...

Bioluminescent Orthotopic Model of Pancreatic Cancer Progression

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%. New therapeutic options are critically needed and depend on improved understanding of pancreatic cancer biology. To better understand the interaction of cancer cells with the pancreatic microenvironment, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression. Luciferase-tagged pancreatic cancer cells are resuspended in Matrigel and delivered into the pancreatic tail during laparotomy. Matrigel solidifies at body temperature to prevent leakage of cancer cells during injection. Primary tumor growth and metastasis to distant organs are monitored following injection of the luciferase substrate luciferin, using in vivo imaging of bioluminescence emission from the cancer cells. In vivo imaging also may be used to track primary tumor recurrence after resection. This orthotopic model is suited to both syngeneic and xenograft models and may be used in pre-clinical trials to investigate the impact of novel anti-cancer therapeutics on the growth of the primary pancreatic tumor and metastasis.

Improved understanding of pancreatic cancer biology is critically needed to enable the development of better therapeutic options to treat pancreatic cancer. To address this need, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression using in vivo bioluminescence imaging.

Modelo ortotópico bioluminiscente de la progresión del cáncer de páncreas

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

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  ...

Minimally Invasive Establishment of Murine Orthotopic Bladder Xenografts

Orthotopic bladder cancer xenografts are the gold standard to study molecular cellular manipulations and new therapeutic agents&#160;in vivo. Suitable cell lines are inoculated either by intravesical instillation (model of nonmuscle invasive growth) or intramural injection into the bladder wall (model of invasive growth). Both procedures are complex and highly time-consuming. Additionally, the superficial model has its shortcomings due to the lack of cell lines that are tumorigenic following instillation. Intramural injection, on the other hand, is marred by the invasiveness of the procedure and the associated morbidity for the host mouse.
With these shortcomings in mind, we modified previous methods to develop a minimally invasive approach for creating orthotopic bladder cancer xenografts. Using ultrasound guidance we have successfully performed percutaneous inoculation of the bladder cancer cell lines UM-UC1, UM-UC3 and UM-UC13 into 50 athymic nude. We have been able to demonstrate that this approach is time efficient, precise and safe. With this technique, initially a space is created under the bladder mucosa with PBS, and tumor cells are then injected into this space in a second step. Tumor growth is monitored at regular intervals with bioluminescence imaging and ultrasound. The average tumor volumes increased steadily in in all but one of our 50 mice over the study period.
In our institution, this novel approach, which allows bladder cancer xenograft inoculation in a minimally-invasive, rapid and highly precise way, has replaced the traditional model.

The established technique to inoculate primary invasive orthotopic bladder cancer xenografts requires laparotomy and mobilization of the bladder. This procedure inflicts significant morbidity on the mice, is technically challenging and time-consuming. We therefore developed a high-precision, percutaneous approach utilizing ultrasound guidance.

Mínimamente Invasiva Establecimiento de murino ortotópico xenoinjertos vejiga

Orthotopic bladder cancer xenografts are the gold standard to study molecular cellular manipulations and new therapeutic agents&#160;in vivo. Suitable ...