Name: a murine orthotopic bladder tumor model and tumor detection system
Uploaded: 2017-01-12T15:00:00.000+00:00
Duration: 6 min 23 s
Description: Watch this Scientific Journal Video about A Murine Orthotopic Bladder Tumor Model and Tumor Detection System at JoVE.com

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

<ol>
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The authors have nothing to disclose.

Os passos mais críticos no protocolo são: 1) manter com sucesso a tumorigenicidade da linha celular; 2) assegurando a secreção PSA mensurável antes da implantação de células tumorais em camundongos; 3) a geração de uma suspensão de uma única célula para o implante, de modo a reduzir a variação no tamanho do tumor; e 4) células instilar a uma velocidade lenta para evitar o refluxo vesico-ureteral, resultando na implantação de células de tumor no rim. Após passagem prolonga...

O objetivo deste método é gerar tumores de bexiga murino ortotópico e monitorar os tumores implantados mais exacta possível, de modo que os ratos sem a implantação do tumor não são pensados ​​para ter sido curado a análise de ponto final. Em geral, o método mostrado irá reduzir a necessidade de grandes números de ratinhos para análise experimental e assegurar uma maior precisão na determinação dos resultados terapêuticos. O desenvolvimento de um modelo ortotópico de câncer é importante, como células tumorais implantando subcutânea não recapitular o ambiente da doença clínica ou permitir o desenvolvimento de estratégias terapêuticas. A arquitectura da bexiga permite a instilação de terapias do cancro da bexiga directamente na bexiga com efeitos sistémicos mínimos. Assim, os modelos animais que recapitulam neste ambiente, tais como um modelo ortotópico, são importantes para avaliar novas terapias. As conclusões tiradas a partir de qualquer experimental set-up são dependenT sobre as limitações do modelo. Diversas técnicas têm sido desenvolvidas para a produção de tumores da bexiga ortotópico em murganhos. Estes dependem danificar a camada de glicosaminoglicano da bexiga, permitindo que as células tumorais a ser implantado. As técnicas utilizadas incluem a electrocauterização, o que resulta em um único ponto de dano na parede da bexiga, conduzindo ao desenvolvimento de tumores em um local na bexiga 1,2. No entanto, a taxa de sucesso da implantação do tumor usando electrocauterização é dependente do operador variando de 10 - 90%, e inclui o risco de que a parede da bexiga será perfurado, levando a desenvolver tumores na cavidade peritoneal. Cauterização química é realizada utilizando nitrato de prata, que danifica a parede da bexiga 3. Da mesma forma, o ácido foi utilizado para danificar a parede da bexiga 4. A tripsina também tem sido utilizado para danificar a bexiga, bem 5. Estes métodos podem resultar no desenvolvimento de mais de um tumor na bexiga.Além disso, existe o perigo de danos graves para a bexiga se os produtos químicos são deixados em contacto com a parede da bexiga por muito tempo. O método desenvolvido por Ninalga et ai. utiliza o poli-L-lisina carregada positivamente (PLL) 6 moléculas para o revestimento da parede da bexiga; isso permite que as células tumorais carregadas negativamente para furar a camada de glicosaminoglicanos da bexiga. Este método resulta geralmente em mais do que um tumor em desenvolvimento na bexiga, mas a implantação do tumor é a 80 - 100% 4,7. Tecnicamente, é também o processo mais fácil de executar. Para assegurar que os tumores que se desenvolvem são bastante uniforme em tamanho, é importante que as células tumorais não são agrupados em grandes aglomerados antes do implante. A fim de avaliar a eficácia terapêutica, é melhor para realizar este estudo em ratos com tumores bastante de tamanho semelhante. Assim, um sistema de detecção de boa que pode quantificar o tamanho do tumor logo após o implante é importante. Várias estratégias têm a abelhaN utilizado para avaliar tumores. Estes incluem a imagiologia por ressonância magnética (MRI) de 8-10, de fluorescência 11, bioluminescência 12,13, ultra-sons 14, e ensaio imunoabsorvente ligado a enzima (ELISA) 15,16. Enquanto ressonância magnética e ultra-som não requerem modificações de células tumorais, existe uma necessidade de equipamentos e agentes de contraste para ressonância magnética sensíveis. Os ensaios fluorescence-, luminescence-, e baseado em ELISA exigir a modificação das células tumorais para expressar proteínas marcadoras que podem ser detectados por estes métodos. Para luminescência, um substrato é necessária para a detecção da actividade de luciferase; Assim, existe um passo adicional e aumento de custo. Ambos luminescência e fluorescência requerem equipamento especializado. Para produzir fluorescência, proteína fluorescente verde (GFP) ciclização, a qual é catalisada por oxigénio molecular, é necessária. Assim, a expressão GFP pode ser variável dentro de uma massa tumoral dependendo do acesso ao oxigênio, tornando este um marcador pouco fiável <sup&gt; 17. Modificação da linha de células de cancro da bexiga murino MB49 a secretar próstata humana Specific Antigen (PSA) 15,16 como um marcador substituto é outra estratégia. Esses marcadores também fornecem um meio alternativo de confirmar a presença do tumor no final da experiência, tornando-os uma alternativa à imuno-histoquímica. Este estudo relata o método PLL de implantação do tumor ortotópico e apresenta uma comparação de sistemas de detecção de tumor, isto é, ELISA, a fluorescência, e ultra-sonografia.

<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 descreve a geração de tumores da bexiga em ratos C57BL / 6J ratinhos fêmea, utilizando a linha celular de cancro da bexiga de murino MB49, que foi modificado para secretar humano Antigénio Específico da Próstata (PSA), e o procedimento para a confirmação da implantação do tumor. Em resumo, os ratos são anestesiados usando drogas injectáveis ​​e são feitos para colocar na posição dorsal. A urina é desocupado da bexiga e 50 ul de poli-L-lisina (PLL) é instilada lentamente a uma taxa de 10 mL / 20 s utilizando um cateter de 24 g IV. É deixado na bexiga durante 20 minutos por tapando o cateter. O cateter é retirado e PLL é desocupado por uma leve pressão sobre a bexiga. Isto é seguido por instilação da linha celular de cancro da bexiga de murino (1 x 10 5 células / 50 ul) a uma taxa de 10 mL / 20 s. O cateter é tapado para evitar a evacuação prematura. Após 1 h, os ratinhos são reavivado com uma droga inversão, e a bexiga está desocupado. A taxa de instilação lenta é importante,uma vez que reduz o refluxo vesico-ureteral, o que pode causar tumores a ocorrer no tracto urinário superior e nos rins. A linha de células deve ser bem re-suspensa para reduzir a aglutinação de células, como esta pode levar à tamanhos irregulares após a implantação do tumor. Esta técnica induz tumores com alta eficiência. O crescimento do tumor é monitorizado pela secreção urinária PSA. monitorização PSA marcador é mais fiável do que o ultra-som ou de imagem de fluorescência para a detecção da presença de tumores na bexiga. Os tumores em ratinhos geralmente atingem um tamanho máximo que afeta negativamente a saúde em cerca de 3-4 semanas, se deixados sem tratamento. Ao monitorizar o crescimento do tumor, é possível diferenciar os ratinhos que estavam curados do que aqueles que não foram implantadas com sucesso com tumores. Com a análise de ponto final, apenas, o último pode ser assumida por engano ter sido curado por terapia.

a secreção de PSA a partir de células MB49 foi encontrada a variar com o meio de crescimento. MB49-PSA é cultivada em meio DMEM, porque isto resulta em aumento da secreção de PSA (Figura 1A). A fim de determinar a sensibilidade do PSA ELISA e PCR em tempo real, os diferentes números de MB49-secretoras de PSA células foram misturadas com células parentais MB49. PSA ELISA detecta um mínimo de 1 x 10 5 células secretoras de PSA / 1 x 10 6 c?...

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 descreve a geração de tumores da bexiga ortotópico de murídeo em Ratinhos C57BL / 6J e a monitorização do crescimento do tumor.

Um modelo de tumor de murino de bexiga ortotópico e Tumor Detection System

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

14.6K visualizações.  National University of Singapore, National University Health System.  Este protocolo descreve a geração de tumores da bexiga ortotópico de murídeo em Ratinhos C57BL / 6J e a monitorização do crescimento do tumor. 

Vídeo: 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 Experimental Modelo de desenvolvimento do tumor original e Peritoneal Metástase<em&gt; via</em&gt; Ortotópico injetar células do ovário Carcinoma

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

Pesquisa do câncer

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.

Um modelo ortotópico de câncer de bexiga 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.

Ressonância magnética avaliação dos tumores de bexiga murino induzida por substância cancerígena

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.

Um modelo de tumor de bexiga ortotópico e Avaliação de Tratamento de sarna intravesical

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.

Cultura, Manipulação e Transplante Ortotópico de Organoides tumorais da bexiga de camundongo

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.

Um modelo de cancro da bexiga para Ortotópico Gene Estudos de entrega

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.

Minimamente Invasiva Estabelecimento de murino ortotópico bexiga Xenoenxertos

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.

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 de Tumor de Bexiga: Um Método para Transplante Ortotópico de Organoides de Bexiga para Bexiga 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

Câncer do trato urinário

Estudos com modelo animal

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.

Modelagem de tumor ortotópico de bexiga em modelo de camundongo: um procedimento para administração intravesical de células cancerígenas na bexiga 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 ...