This work was funded by a grant from the National Medical Research Council of Singapore (NMRC/CIRG/1335/2012) awarded to Professor Kesavan Esuvaranathan.

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</ol>

The authors have nothing to disclose.

Los pasos más críticos en el protocolo son 1) el mantenimiento con éxito la tumorigenicidad de la línea celular; 2) asegurar la secreción de PSA medible antes de la implantación de células tumorales en ratones; 3) la generación de una suspensión de una sola célula para la implantación con el fin de reducir la variación en el tamaño del tumor; y 4) infundir células a una velocidad lenta para prevenir el reflujo vesico-ureteral, lo que resulta en la implantación de células tumorales en el riñón. <p c...

El objetivo de este método es generar tumores de vejiga murino ortotópico y para controlar los tumores implantados con la mayor precisión posible, de manera que los ratones sin la implantación del tumor no se cree que se han curado a análisis de punto final. En general, el método que se muestra reducirá la necesidad de un gran número de ratones para el análisis experimental y garantizar una mayor precisión en la determinación de los resultados terapéuticos. El desarrollo de un modelo ortotópico de cáncer es importante, ya que la implantación de las células tumorales por vía subcutánea no recapitula el medio ambiente de la enfermedad clínica o permiten el desarrollo de estrategias terapéuticas. La arquitectura de la vejiga permite la instilación de las terapias de cáncer de vejiga directamente en la vejiga con efectos sistémicos mínimos. Por lo tanto, los modelos animales que recapitular este entorno, tales como un modelo ortotópico, son importantes para evaluar nuevas terapias. Las conclusiones extraídas de cualquier montaje experimental se dependent sobre las limitaciones del modelo. Varias técnicas han sido desarrolladas para la producción de los tumores de vejiga ortotópico en ratones. Estos dependen de dañar la capa de glicosaminoglicano de la vejiga, lo que permite las células tumorales para ser implantado. Las técnicas utilizadas incluyen la electrocauterización, que resulta en un único punto de daño en la pared de la vejiga, lo que lleva al desarrollo de tumores en un sitio en la vejiga 1,2. Sin embargo, la tasa de éxito de la implantación del tumor usando electrocauterización es dependiente del operador que varía de 10 - 90%, y que incluye el peligro de que la pared de la vejiga se punza, lo que lleva a los tumores en desarrollo en la cavidad peritoneal. Cauterización química se lleva a cabo utilizando nitrato de plata, lo que daña la pared de la vejiga 3. Del mismo modo, el ácido se ha utilizado para dañar la pared de la vejiga 4. La tripsina también se ha utilizado para dañar la vejiga, así 5. Estos métodos pueden resultar en el desarrollo de más de un tumor en la vejiga.Además, existe el peligro de un daño severo a la vejiga si los productos químicos se dejan en contacto con la pared de la vejiga durante demasiado tiempo. El método desarrollado por Ninalga et al. utiliza el poli-L-lisina cargada positivamente (PLL) 6 moléculas para recubrir la pared de la vejiga; esto permite que las células tumorales con carga negativa a pegarse a la capa de glicosaminoglicano de la vejiga. Este método da como resultado en general en más de un tumor en desarrollo en la vejiga, pero la implantación del tumor es a 80 - 100% de 4,7. Técnicamente, también es el procedimiento más fácil de realizar. Para asegurarse de que los tumores que se desarrollan son bastante uniforme en tamaño, es importante que las células tumorales no se agrupan en grupos grandes antes de la implantación. Con el fin de evaluar la eficacia terapéutica, lo mejor es llevar a cabo este estudio en ratones con tumores bastante de tamaño similar. Por lo tanto, un buen sistema de detección que puede cuantificar el tamaño del tumor pronto después de la implantación es importante. Varias estrategias han de abejasn utiliza para evaluar tumores. Estos incluyen imágenes por resonancia magnética (IRM) 8-10, 11 de fluorescencia, bioluminiscencia 12,13, ultrasonido 14, y el ensayo de inmunoabsorción ligado a enzimas (ELISA) 15,16. Mientras que la RM y la ecografía no requieren modificaciones de las células tumorales, hay una necesidad de equipos y agentes de contraste para resonancia magnética sensibles. Los ensayos por fluorescencia, luminiscencia, y basados ​​en ELISA requieren modificación de las células tumorales para expresar proteínas marcadoras que se pueden detectar por estos métodos. Por luminiscencia, se requiere un sustrato para la detección de la actividad de la luciferasa; Por lo tanto, hay un paso adicional y aumento del coste. Tanto la luminiscencia y fluorescencia requieren equipo especializado. Para producir fluorescencia, proteína fluorescente verde (GFP) de ciclación, que es catalizada por el oxígeno molecular, se requiere. Por lo tanto, la expresión de GFP puede ser variable dentro de una masa tumoral en función de acceso al oxígeno, haciendo de este un marcador más fiable <sup&gt; 17. La modificación de la línea celular de cáncer de vejiga murino MB49 para secretar próstata humana Antígeno Específico (PSA) 15,16 como un marcador sustituto es otra estrategia. Estos marcadores también proporcionan un medio alternativo para confirmar la presencia del tumor en la terminación del experimento, por lo que una alternativa a la inmunohistoquímica. Este estudio reporta el método PLL de la implantación del tumor ortotópico y presenta una comparación de los sistemas de detección de tumores, a saber, ELISA, fluorescencia, y las imágenes por ultrasonido.

<table><tbody><tr><td>MB49-PSA cells</td><td>N/A</td><td>N/A</td><td>ref (Wu QH, 2004)</td></tr><tr><td>RPMI 1640 media</td><td>HyClone</td><td>SH30027.01&amp;nbsp;</td><td></td></tr><tr><td>Dulbecco&#039;s Modified Eagle&#039;s Medium (DMEM) media</td><td>Biowest</td><td>L0102</td><td></td></tr><tr><td>Fetal Bovine Serum (FBS) South American</td><td>Biowest</td><td>S1810</td><td></td></tr><tr><td>Fetal Bovine Serum (FBS) South American, Premium</td><td>Biowest</td><td>S181B</td><td></td></tr><tr><td>Fetal Bovine Serum (FBS)</td><td>HyClone</td><td>SH30088.03&amp;nbsp;</td><td></td></tr><tr><td>L-glutamine</td><td>Biowest</td><td>X0550</td><td></td></tr><tr><td>Penicillin-Streptomycin</td><td>Biowest</td><td>L0022</td><td></td></tr><tr><td>Hygromycin B</td><td>Invitrogen</td><td>10687-010</td><td></td></tr><tr><td>free PSA (Human) ELISA kit</td><td>Abnova</td><td>KA0209</td><td></td></tr><tr><td>TRIzol Reagent for RNA extraction&amp;nbsp;</td><td>Ambion</td><td>15596026</td><td></td></tr><tr><td>High Capacity cDNA Reverse Transcription Kit with RNase Inhibitor</td><td>Applied Biosystems</td><td>4374967</td><td></td></tr><tr><td>TaqMan Universal PCR Master Mix</td><td>Applied Biosystems</td><td>4304437</td><td></td></tr><tr><td>TaqMan Gene Expression Assay &amp;ndash; Mouse Actb</td><td>Applied Biosystems</td><td>4331182</td><td>Mm00607939_s1</td></tr><tr><td>TaqMan Gene Expression Assay &amp;ndash; Human KLK3</td><td>Applied Biosystems</td><td>4331182</td><td>Hs00426859_g1</td></tr><tr><td>C57BL/6J female mice</td><td>In Vivos</td><td></td><td>4 - 6 wk old</td></tr><tr><td>Anesthesia (75 mg/kg Ketamine and 1 mg/kg Medetomidine)</td><td>Local pharmacy</td><td></td><td></td></tr><tr><td>Reversal drug (1 mg/kg Atipamezole)</td><td>Local pharmacy</td><td></td><td></td></tr><tr><td>Ear punch</td><td>Electron Microscopy Sciences</td><td>72893-01</td><td></td></tr><tr><td>Hartmann&#039;s solution or Compound sodium lactate</td><td>B Braun</td><td></td><td></td></tr><tr><td>Ophthalmic ointment - Duratears sterile ocular lubricant ointment</td><td>Alcon</td><td></td><td></td></tr><tr><td>Heat pack - HotHands handwarmers</td><td>Heatmax Inc</td><td></td><td></td></tr><tr><td>Introcan Certo IV catheter</td><td>B Braun</td><td>4251300</td><td>24 G x 3/4&amp;Prime;</td></tr><tr><td>Aquagel Lubricating jelly</td><td>Local pharmacy</td><td></td><td></td></tr><tr><td>Poly-L-lysine solution, 0.01%,</td><td>Sigma</td><td>P4707</td><td></td></tr><tr><td>cOmplete, Mini, EDTA-free Protease Inhibitor Cocktail</td><td>Roche</td><td>4693159001</td><td></td></tr><tr><td>Quantichrom Creatinine Assay Kit</td><td>BioAssay Systems</td><td>DICT-50&amp;nbsp;</td><td></td></tr><tr><td>Fluorescent dye - VivoTrack 680</td><td>Perkin Elmer</td><td>NEV12000&amp;nbsp;</td><td></td></tr><tr><td>RNAlater-ICE Frozen Tissue Transition Solution</td><td>Ambion</td><td>4427575</td><td></td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Equipment and Software&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>7500 Realtime PCR System</td><td>Applied Biosystems</td><td></td><td></td></tr><tr><td>7500 Software v2.3</td><td>Applied Biosystems</td><td></td><td></td></tr><tr><td>Metabolic Cage</td><td>Tecniplast&amp;nbsp;</td><td>Vertical type rack for 12 cages</td><td></td></tr><tr><td>BD FACSCanto I system&amp;nbsp;</td><td>BD Biosciences</td><td></td><td></td></tr><tr><td>BD FACSDiva software v7</td><td>BD Biosciences</td><td></td><td></td></tr><tr><td>IVIS SpectrumCT &lt;em&gt;in vivo&lt;/em&gt; imaging system&amp;nbsp;</td><td>Caliper Life Sciences&amp;nbsp;</td><td></td><td></td></tr><tr><td>Living Image Software v3.1</td><td>Caliper Life Sciences&amp;nbsp;</td><td></td><td></td></tr><tr><td>Vevo 2100 imaging system&amp;nbsp;</td><td>VisualSonics&amp;nbsp;</td><td></td><td></td></tr></tbody></table>

a murine orthotopic bladder tumor model and tumor detection system

Este protocolo describe la generación de tumores de vejiga en ratones C57BL / 6J hembra utilizando la línea celular de cáncer de vejiga murino MB49, que ha sido modificado para secretar de próstata humano de antígeno específico (PSA), y el procedimiento para la confirmación de la implantación del tumor. En resumen, los ratones se anestesiaron usando drogas inyectables y están hechos para poner en la posición dorsal. La orina está desocupado desde la vejiga y 50 l de poli-L-lisina (PLL) se instila lentamente a una velocidad de 10 l / s 20 utilizando un catéter 24 G IV. Se deja en la vejiga durante 20 min por tapando el catéter. Se retira el catéter y el PLL está desocupado por una suave presión sobre la vejiga. Esto es seguido por la instilación de la línea celular de cáncer de vejiga murino (1 x 10 5 células / 50 l) a una velocidad de 10 l / 20 s. El catéter se tapó para evitar la evacuación prematura. Después de 1 h, los ratones se revivieron con un fármaco de reversión, y la vejiga esté desocupado. La tasa de instilación lenta es importante,ya que reduce el reflujo vesico-ureteral, lo que puede hacer que los tumores se producen en el tracto urinario superior y en los riñones. La línea celular debe estar bien volvió a suspenderse para reducir la aglutinación de las células, ya que esto puede conducir a tamaños irregulares después de la implantación del tumor. Esta técnica induce tumores con alta eficiencia. El crecimiento tumoral se controló mediante la secreción de PSA urinaria. monitoreo marcador PSA es más fiable que el ultrasonido o imágenes de fluorescencia para la detección de la presencia de tumores en la vejiga. Los tumores en los ratones generalmente alcanzan un tamaño máximo que repercute negativamente en la salud por cerca de 3 - 4 semanas si no se tratan. Mediante el control de crecimiento del tumor, es posible diferenciar los ratones que fueron curados de las que no fueron implantados con éxito con los tumores. Con el análisis de punto final únicamente, el último puede ser erróneamente supone que se han curado mediante terapia.

se encontró la secreción de PSA a partir de células MB49 a variar con el medio de crecimiento. MB49-PSA se cultiva en medios DMEM porque esto da lugar a aumento de la secreción de PSA (Figura 1A). Con el fin de determinar la sensibilidad de la PSA ELISA y PCR en tiempo real, diferentes números de células MB49-PSA secretoras se mezclaron con células parentales MB49. PSA ELISA detecta un mínimo de 1 x 10 5 células secretoras de PSA / 1 x 10 6...

Watch this Scientific Journal Video about A Murine Orthotopic Bladder Tumor Model and Tumor Detection System at JoVE.com

A Murine Orthotopic Bladder Tumor Model and Tumor Detection System

Este protocolo describe la generación de tumores de vejiga ortotópico murino en ratones C57BL / 6J hembra y el seguimiento del crecimiento del tumor.

Un Modelo de tumor y el tumor murino ortotópico de vejiga Sistema de Detección

This protocol describes the generation of bladder tumors in female C57BL/6J mice using the murine bladder cancer cell line MB49, which has been modified ...

a-murine-orthotopic-bladder-tumor-model-and-tumor-detection-system

Research

JoVE Journal

Cancer Research

14.7K vistas.  National University of Singapore, National University Health System.  Este protocolo describe la generación de tumores de vejiga ortotópico murino en ratones C57BL / 6J hembra y el seguimiento del crecimiento del tumor. 

Video: A Murine Orthotopic Bladder Tumor Model and Tumor Detection System

Murine Experimental Model of Original Tumor Development and Peritoneal Metastasis via Orthotopic Inoculation with Ovarian Carcinoma Cells

Epithelial ovarian carcinoma (EOC) is associated with a poor prognosis because it shows peritoneal dissemination. To improve the prognosis, it is important to control peritoneal dissemination. However, it is still unclear how tumor cells detach from primary lesions and attach to the mesothelium. The establishment of an appropriate animal model is needed to gain an understanding of the mechanism of peritoneal dissemination in vivo. In the current study, we introduce the process from the local injection of EOC cells into the murine ovarian surface to the development of metastasis, including the peritoneum and distant organs. Female nude mice (BALB/c nu/nu) at 8 weeks of age were used. Under a microscopic field of view, EOC cells (1 x 105 cells/&#181;l of medium-extracellular matrix (ECM)-based hydrogel/unilateral ovary/mouse) were injected into murine ovaries through a retroperitoneal approach from the dorsal flank. This proposed method is a less invasive procedure for the mouse and minimizes damage to the ovary. Here, we describe the methodological steps in the development of the original and metastatic tumor formation of EOC.

Murine Experimental Model of Original Tumor Development and Peritoneal Metastasis via Orthotopic Inoculation with Ovarian Carcinoma Cells

To clarify the multistep process of peritoneal dissemination of ovarian carcinoma, the current study presents a murine experimental model of original tumor development and peritoneal metastasis via orthotopic inoculation with these tumor cells.

Murino modelo experimental de desarrollo del tumor original y la metástasis peritoneal<em&gt; vía</em&gt; Ortotópico La inoculación con células de carcinoma de ovario

Epithelial ovarian carcinoma (EOC) is associated with a poor prognosis because it shows peritoneal dissemination. To improve the prognosis, it is ...

Investigación

Investigación sobre el cáncer

An Orthotopic Model of Murine Bladder Cancer

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

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

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 ...

Medicina

Magnetic Resonance Imaging Assessment of Carcinogen-induced Murine Bladder Tumors

Murine bladder tumor models are critical for the evaluation of new therapeutic options. Bladder tumors induced with the N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) carcinogen are advantageous over cell line-based models because they closely replicate the genomic profiles of human tumors, and, unlike cell models and xenografts, they provide a good opportunity for the study of immunotherapies. However, bladder tumor generation is heterogeneous; therefore, an accurate assessment of tumor burden is needed before randomization to experimental treatment. Described here is a BBN mouse model and protocol to evaluate bladder cancer tumor burden in vivo using a fast and reliable magnetic resonance (MR) sequence (true FISP). This method is simple and reliable because, unlike ultrasound, MR is operator-independent and allows for the straightforward post-acquisition image processing and review. Using axial images of the bladder, analysis of regions of interest along the bladder wall and tumor allow for the calculation of bladder wall and tumor area. This measurement correlates with ex vivo bladder weight (rs= 0.37, p = 0.009) and tumor stage (p = 0.0003). In conclusion, BBN generates heterogeneous tumors that are ideal for evaluation of immunotherapies, and MRI can quickly and reliably assess tumor burden prior to randomization to experimental treatment arms.

Murine bladder tumors are induced with the N-butyl-N-(4-hydroxybutyl) nitrosamine carcinogen (BBN). Bladder tumor generation is heterogeneous; therefore, an accurate assessment of tumor burden is needed before randomization to experimental treatment. Here we present a fast, reliable MRI protocol to assess tumor size and stage.

Evaluación de la proyección de imagen de resonancia magnética de los tumores vesicales murina inducida por carcinógenos

Murine bladder tumor models are critical for the evaluation of new therapeutic options. Bladder tumors induced with the N-butyl-N-(4-hydroxybutyl) ...

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 ...

Culture, Manipulation, and Orthotopic Transplantation of Mouse Bladder Tumor Organoids

The development of advanced tumor models has long been encouraged because current cancer models have shown limitations such as lack of three-dimensional (3D) tumor architecture and low relevance to human cancer. Researchers have recently developed a 3D in vitro cancer model referred to as tumor organoids that can mimic the characteristics of a native tumor in a culture dish. Here, experimental procedures are described in detail for the establishment of bladder tumor organoids from a carcinogen-induced murine bladder tumor, including culture, passage, and maintenance of the resulting 3D tumor organoids in vitro. In addition, protocols to manipulate the established bladder tumor organoid lines for genetic engineering using lentivirus-mediated transduction are described, including optimized conditions for the efficient introduction of new genetic elements into tumor organoids. Finally, the procedure for orthotopic transplantation of bladder tumor organoids into the wall of the murine bladder for further analysis is laid out. The methods described in this article can facilitate the establishment of an in vitro model for bladder cancer for the development of better therapeutic options.

This protocol provides detailed experimental steps to establish a three-dimensional in vitro culture of bladder tumor organoids derived from carcinogen-induced murine bladder cancer. Culture methods including passaging, genetic engineering, and orthotopic transplantation of tumor organoids are described.

Cultivo, manipulación y trasplante ortotópico de organoides tumorales de la vejiga de ratón

The development of advanced tumor models has long been encouraged because current cancer models have shown limitations such as lack of three-dimensional ...

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 ...

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 ...

Establishing a Murine Superficial Bladder Cancer Model via an Intravesical Cell Administration Technique

This study presents an innovative method for establishing an orthotopic murine bladder tumor model with high efficiency and precise tumor localization. After anesthetizing female C57BL/6J mice in a supine position, a 24 G intravenous needle is inserted into the bladder to evacuate its contents. A 34 G dispensing needle is then introduced through the catheter, rotated five times to create a focal injury to the bladder dome mucosa, and subsequently removed. The MB49 cell suspension is aspirated and connected to a 30 G dispensing needle, which is inserted into the bladder via the catheter. Tumor cells are injected submucosally into the bladder under pressure. This technique results in minimal trauma to the mice, a high tumor take rate, and a fixed tumor location. It is characterized by simplicity and excellent reproducibility. This model provides an ideal experimental platform for developing intravesical therapies for bladder cancer, facilitating the advancement and optimization of treatment strategies for this malignancy.

This protocol describes a unique method for constructing an orthotopic model of superficial bladder cancer.

Establecimiento de un modelo de cáncer de vejiga superficial murino mediante una técnica de administración de células intravesicales

This study presents an innovative method for establishing an orthotopic murine bladder tumor model with high efficiency and precise tumor localization. ...

Bladder Tumor Organoid Model: A Method for Orthotopic Transplantation of Bladder Organoids into Recipient Murine Bladder

Source: Kim, Y. et al. <a href="https://www.jove.com/video/60469" target="_blank">Culture, Manipulation, and Orthotopic Transplantation of Mouse Bladder Tumor Organoids.</a>&#160;J. Vis. Exp.&#160;(2020)
In this video, we describe the orthotopic transplantation of cultured bladder tumor organoids into the murine bladder to study the role of the organ environment on developing tumors.

Modelo de organoide tumoral vesical: un método para el trasplante ortotópico de organoides vesicales en vejiga murina receptora

Source: Kim, Y. et al. Culture, Manipulation, and Orthotopic Transplantation of Mouse Bladder Tumor Organoids.&#160;J. Vis. Exp.&#160;(2020)
In this ...

JoVE Encyclopedia of Experiments

Urinary Tract Cancer

In vivo studies

Modeling Orthotopic Bladder Tumor in Mouse Model: A Procedure for Intravesical Administration of Cancer Cells into Murine Bladder

Source: Tham, S. M. et al. <a href="https://www.jove.com/video/55078" target="_blank">A Murine Orthotopic Bladder Tumor Model and Tumor Detection System.</a>&#160;J. Vis. Exp.&#160;(2017)
In this video, we describe the intravesical procedure of delivering cancer cells into the bladder of a mouse model. This procedure helps generate an orthotopic bladder tumor model that can be used to develop therapeutic strategies.

Modelado de tumor ortotópico de vejiga en modelo de ratón: un procedimiento para la administración intravesical de células cancerosas en la vejiga murina

Source: Tham, S. M. et al. A Murine Orthotopic Bladder Tumor Model and Tumor Detection System.&#160;J. Vis. Exp.&#160;(2017)
In this video, we describe ...