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.

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The authors have nothing to disclose.

الخطوات الأكثر أهمية في البروتوكول هي 1) الحفاظ بنجاح tumorigenicity من خط الخلية. 2) ضمان إفراز دعم البرامج والإدارة للقياس قبل زرع الخلايا السرطانية في الفئران. 3) توليد تعليق خلية واحدة لزرع ذلك للحد من التباين في حجم الورم. و4) غرس الخلايا بمعدل بطيء لمنع المثاني الحالبي، ا?...

والهدف من هذه الطريقة هو توليد أورام المثانة مثلي الفئران ورصد الأورام مزروع بأكبر قدر ممكن، حتى لا يعتقد أن الفئران دون زرع الورم قد تم علاجه في تحليل نقطة النهاية. وعموما، فإن طريقة أظهرت سوف يقلل من الحاجة لأعداد كبيرة من الفئران لتحليل تجريبي وضمان المزيد من الدقة في تحديد النتائج العلاجية. تطوير نموذج مثلي لسرطان مهمة، كما زرع خلايا الورم تحت الجلد لا ألخص البيئة من المرض السريري أو تمكين وضع استراتيجيات علاجية. بنية المثانة يسمح للتقطير من علاجات سرطان المثانة مباشرة في المثانة مع تأثيرات جهازية الحد الأدنى. وبالتالي، نماذج حيوانية أن ألخص هذه البيئة، مثل نموذج مثلي، هي مهمة لتقييم علاجات جديدة. وdependen الاستنتاجات المستخلصة من أي انشاء التجريبيةر على القيود المفروضة على النموذج. وقد تم تطوير العديد من التقنيات لإنتاج أورام المثانة مثلي في الفئران. هذه تعتمد على إتلاف طبقة جلايكان المثانة، مما يتيح الخلايا السرطانية ليكون مزروع. وتشمل التقنيات المستخدمة كهربي، مما يؤدي الى نقطة واحدة من الضرر في جدار المثانة، مما يؤدي إلى نمو الورم في موقع واحد في المثانة 1،2. ومع ذلك، فإن نسبة نجاح زرع الورم باستخدام كهربي يعتمد مشغل متفاوتة 10-90٪، ويشمل الخطر الذي جدار المثانة سيتم ثقب، مما يؤدي إلى أورام النامية في التجويف البريتوني. يتم تنفيذ الكي الكيميائي باستخدام نترات الفضة، الامر الذي يسيء الى جدار المثانة 3. وبالمثل، وقد استخدم حمض إلى تلف جدار المثانة 4. كما تم استخدام التربسين إلى تلف المثانة وكذلك 5. قد ينتج عن هذه الأساليب في تطوير الورم أكثر من واحد في المثانة.وعلاوة على ذلك، هناك خطر حدوث أضرار بالغة في المثانة إذا ترك المواد الكيميائية في اتصال مع جدار المثانة لفترة طويلة جدا. الطريقة التي وضعتها Ninalga وآخرون. يستخدم موجبة بولي-L-ليسين (PLL) 6 الجزيئات إلى طبقة جدار المثانة. وهذا يتيح لخلايا الورم المشحونة سلبا على التمسك طبقة جلايكان المثانة. هذه الطريقة عموما النتائج في الورم أكثر من واحد النامية في المثانة، ولكن الورم غرس هو في 80-100٪ 4،7. من الناحية الفنية، بل هو أيضا أسهل طريقة لأداء. لضمان أن الأورام التي تنشأ هي إلى حد ما حتى في الحجم، من المهم أن الخلايا السرطانية لا تجميعها في كتل كبيرة قبل الزرع. من أجل تقييم فعالية العلاج، فمن الأفضل لأداء هذه الدراسة على الفئران بأورام مماثلة الحجم إلى حد ما. وهكذا، فإن نظام الكشف الجيدة التي يمكن قياس حجم الورم قريبا بعد زرع مهم. العديد من الاستراتيجيات لها النحلن المستخدمة لتقييم الأورام. وتشمل هذه التصوير بالرنين المغناطيسي (MRI) 10/08، مضان 11، تلألؤ بيولوجي 12،13، الموجات فوق الصوتية 14، وانزيم مرتبط المناعي فحص (ELISA) 15،16. بينما التصوير بالرنين المغناطيسي والموجات فوق الصوتية لا تتطلب إدخال تعديلات على الخلايا السرطانية، وهناك حاجة إلى معدات والتباين وكلاء حساسة للتصوير بالرنين المغناطيسي. تتطلب فحوصات fluorescence-، luminescence-، واستنادا ELISA-تعديل الخلايا السرطانية للتعبير عن البروتينات علامة التي يمكن الكشف عنها من قبل هذه الأساليب. لالتلألؤ، مطلوب ركيزة للكشف عن luciferase النشاط. وبالتالي، هناك خطوة إضافية وزيادة تكلفة. كل من التألق ومضان يتطلب معدات متخصصة. لإنتاج مضان، الأخضر بروتين فلوري (GFP) cyclization، الذي يحفزه الأكسجين الجزيئي، مطلوب. وبالتالي، قد يكون التعبير GFP متغير داخل كتلة الورم اعتمادا على الحصول على الأكسجين، مما يجعل هذا علامة لا يمكن الاعتماد عليها بدلا <sup&gt; 17. تعديل الفئران خط خلية سرطان المثانة MB49 لإفراز البروستاتا البشرية مستضد النوعي (PSA) 15،16 كمؤشر بديل هو استراتيجية أخرى. كما توفر هذه العلامات وسيلة بديلة لتأكيد وجود ورم في إنهاء التجربة، مما يجعلها بديلا لالمناعية. تعرض هذه الدراسة المنهج PLL زرع ورم مثلي وتقدم مقارنة بين أنظمة الكشف عن الورم، وهي ELISA، مضان، والتصوير بالموجات فوق الصوتية.

<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

يصف هذا البروتوكول توليد أورام المثانة لدى الفئران C57BL / 6J الإناث باستخدام الفئران خط خلية سرطان المثانة MB49، الذي تم تعديله لإفراز البروستاتا البشرية محددة مستضد (PSA)، وإجراءات التأكد من زرع الورم. باختصار، يتم تخدير الفئران باستخدام المخدرات عن طريق الحقن ومصنوعة لوضع في موقف الظهرية. وأخلى البول من المثانة و 50 ميكرولتر من بولي-L-ليسين (PLL) وتغرس ببطء بمعدل 10 ميكرولتر / 20 ق باستخدام القسطرة 24 G الرابع. فإنه لا يتبقى في المثانة لمدة 20 دقيقة قبل stoppering القسطرة. تتم إزالة القسطرة وجلت PLL بواسطة ضغط لطيف على المثانة. ويعقب ذلك تقطير خط خلية سرطان المثانة الفئران (1 × 10 5 خلية / 50 ميكرولتر) بمعدل 10 ميكرولتر / 20 ق. ومقفول القسطرة لمنع إخلاء سابق لأوانه. بعد 1 ساعة، وإحياء الفئران بعقار انعكاس، وأخلت المثانة. معدل تقطير بطيء مهم،كما أنه يقلل من الجزر المثاني الحالبي، والتي يمكن أن تسبب الأورام تحدث في المسالك البولية العلوي وفي الكلى. خط الخلية يجب أيضا إعادة وقف التنفيذ للحد من التثاقل من الخلايا، وهذا يمكن أن يؤدي إلى أحجام الأورام متفاوتة بعد الزرع. هذه التقنية تحرض الأورام بكفاءة عالية. ويتم رصد نمو الورم عن طريق إفراز دعم البرامج والإدارة البولية. مراقبة PSA علامة هو أكثر موثوقية من الموجات فوق الصوتية أو التصوير مضان للكشف عن وجود أورام في المثانة. الأورام في الفئران تصل عموما حجم أقصى يؤثر سلبا على الصحة بنحو 3-4 أسابيع إذا تركت دون علاج. من خلال رصد نمو الورم، فمن الممكن أن نفرق الفئران التي تم علاجها من تلك التي لم تزرع بنجاح مع الأورام. مع تحليل نقطة النهاية الوحيد، وهذا الأخير يمكن افتراض خاطئ قد تم علاجه عن طريق العلاج.

تم العثور على إفراز دعم البرامج والإدارة من خلايا MB49 أن تختلف مع وسائل الإعلام نمو. يزرع MB49-PSA في وسائل الاعلام DMEM لأن هذا يؤدي إلى زيادة إفراز PSA (الشكل 1A). من أجل تحديد حساسية دعم البرامج والإدارة وإليسا في الوقت الحقيقي PCR، وتباينت أعداد مخت...

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

يصف هذا البروتوكول جيل من الفئران أورام المثانة مثلي في الإناث C57BL / 6J الفئران ورصد نمو الورم.

والفئران مثلي المثانة نموذج ورم والورم نظام لكشف

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

Universidad San Sebastian

Research

JoVE Journal

Cancer Research

14.6K Views.  National University of Singapore, National University Health System.  يصف هذا البروتوكول جيل من الفئران أورام المثانة مثلي في الإناث C57BL / 6J الفئران ورصد نمو الورم. 

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.

فأري موديل التجريبية التنمية الورم الأصلي والبريتوني ورم خبيث<em&gt; عبر</em&gt; مثلي تلقيح مع خلايا المبيض سرطان

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

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.

نموذج مثلي من سرطان المثانة الفئران

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

Medicine

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.

تقييم التصوير بالرنين المغناطيسي لاورام المثانة مورين المستحثة مسرطن

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.

ورم في المثانة Orthotopic النموذجي وتقييم المعالجة سارنا داخل المثانة

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.

الثقافة والتلاعب وزرع تقويم العظام من ورم المثانة الماوس Organoids

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.

وسرطان المثانة نموذج مثلي للدراسات توصيل الجينات

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.

مؤسسة المناظير الفئران مثلي المثانة Xenografts

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.bdigitaluss.remotexs.co/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.

النموذج العضوي لورم المثانة: طريقة لزرع عضيات المثانة في مثانة الفئران المتلقية

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.bdigitaluss.remotexs.co/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.

نمذجة ورم المثانة التقويمي في نموذج الفأر: إجراء للإعطاء داخل المثانة للخلايا السرطانية في مثانة الفئران

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 ...