鼠标膀胱壁注射液
Summary
小鼠膀胱壁注射液是一种有用的方法原位研究膀胱干细胞和癌症生物学。这种微妙的显微外科方法可以掌握仔细的技术和实践。
Abstract
小鼠膀胱壁注射原位研究，包括干细胞，平滑肌，和癌症生物学膀胱现象，是一种有用的技术。开始前注射，手术区必须用肥皂和水和消毒液清洗。在使用前和每个动物之间的手术设备必须消毒。每个鼠标放在吸入异氟醚麻醉下（2-5％的诱导，维持1-3％）和膀胱暴露中线腹部切口，用剪刀。如果膀胱满了，这是部分之间的两根手指轻轻挤压解压。 intramurally利益的细胞悬液注入使用29或30号针头和注射器1毫升或更小的圆顶膀胱壁。伤口闭合伤口剪辑，并允许恢复变暖垫鼠标。膀胱壁注射液是一种微妙的显微外科技术，可以掌握与实践。
Protocol
1。鼠标膀胱壁注射液小鼠品系，年龄和性别的选择是出于实验需要。我们用8至12周龄小鼠，因为这是一个窗口前的免疫成熟到衰老。作为一般指引，老鼠应该到达至少一个星期前，实验操作，以避免应力引起的混杂因素。 用肥皂和水清洁手术台面。 CIDE挥笔或杀菌湿巾擦拭手术台面。 前高压灭菌干净的手术器械，手术中使用。 此外，热珠灭菌消…
Discussion
小鼠膀胱壁注射允许为特定的细胞植入到膀胱壁的定义领域。这项技术已广泛应用，为膀胱癌，平滑肌，和干细胞生物学的小鼠模型。根据定义，小鼠膀胱壁注射液有利于引入膀胱癌或干细胞原位位置，所以可以在一个有关生理解剖背景下的研究，他们的生长和分化。事实上，Dinney等。首次描述了这种技术，以研究在1裸鼠人膀胱癌裸鼠移植瘤原位。
虽然一直在膀胱?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
我们非常感谢支持从斯坦福大学的儿科研究基金和K08DK087895 – 01的NIDDK试点授予。
Materials
| Name of the reagent | Company | Catalogue number |
| Isoflurane (Aerrane) | Baxter | NDC 10019-773-60 |
| 29 or 30 gauge needles | Hamilton | 7803-06 or 7803-07 |
| 100 microliter syringe | Hamilton | 7656-01 |
References
- Dinney, C. P., Fishbeck, R., Singh, R. K., Eve, B., Pathak, S., Brown, N., Xie, B., Fan, D., Bucana, C., Fidler, I. J., Killion, J. J. Isolation and Characterization of Metastatic Variants from Human Transitional Cell Carcinoma Passaged by Orthotopic Implantation in Athymic Nude Mice. The Journal of Urology. 154, 1532-1538 (1995).
- Singh, A. V., Franke, A. A., Blackburn, G. L., Zhou, J. Soy Phytochemicals Prevent Orthotopic Growth and Metastasis of Bladder Cancer in Mice by Alterations of Cancer Cell Proliferation and Apoptosis and Tumor Angiogenesis. Cancer Res. 66, 1851-1858 (2006).
- Yanagiuchi, A., Miyake, H., Nomi, M., Takenaka, A., Fujisawa, M. Modulation of the Microenvironment by Growth Factors Regulates the In Vivo Growth of Skeletal Myoblasts. BJU International. 103, 1569-1573 (2009).
- Miyake, H., Hara, I., Yamanaka, K., Gohji, K., Arakawa, S., Kamidono, S. Overexpression of Bcl-2 Enhances Metastatic Potential of Human Bladder Cancer Cells. British Journal of Cancer. 79, 1651-1656 (1999).
- Chancellor, M. B., Yokoyama, T., Tirney, S., Mattes, C. E., Ozawa, H., Yoshimura, N., Groat, W. C. d. e., Huard, J. Preliminary Results of Myoblast Injection into the Urethra and Bladder Wall: a Possible method for the Treatment of Stress Urinary Incontinence and Impaired Detrusor Contractility. Neurourol Urodyn. 19, 279-287 (2000).
- Dinney, C. P., Tanguay, S., Bucana, C. D., Eve, B. Y., Fidler, I. J. Intravesical Liposomal Muramyl Tripeptide Phosphatidylethanolamine Treatment of Human Bladder Carcinoma Growing in Nude Mice. J Interferon Cytokine Res. 15, 585-592 (1995).
- Mohamedali, K. A., Kedary, D., Sweeney, P., Kamaty, A., Davisy, D. W., Evey, B. Y., Huangy, S., Thorpez, P. E., Dinney, C. P., Rosenblum, M. G. The Vascular-targeting Fusion toxin VEGF121/rGel Inhibits the Growth of Orthotopic Human Bladder Carcinoma Tumors. Neoplasia. 7, 912-920 (2005).
- Slaton, J. W., Perrotte, P., Inoue, K., Dinney, C. P., Fidler, I. J. Interferon-α-mediated Down-Regulation of Angiogenesis-related Genes and Therapy of Bladder Cancer Are Dependent on Optimization of Biological Dose and Schedule. Clinical Cancer Research. 5, 2726-2734 (1999).
- Yokoyama, T., Huard, J., Pruchnic, R., Yoshimura, N., Qu, Z., Cao, B., De Groat, W. C., Kumon, H., Chancellor, M. B. Muscle-Derived Cell Transplantation and Differentiation into Lower Urinary Tract Smooth Muscle. Urology. 57, 826-831 (2001).
- Chan, E., Patel, A., Heston, W., Larchian, W. Mouse Orthotopic Models for Bladder Cancer Research. BJU International. 104, 1286-1291 (1988).
- Russell, P. J., Jelbart, M., Wills, E., Singh, S., Wass, J., Witherspoon, J., Raghavan, D. Establishment and characterization of a new human bladder cancer cell line showing features of squamous and glandular differentiation. Int. J. Cancer. 41, 74-74 (1988).
- Ahlering, T. E., Dubeau, L., Jones, P. A. A new in vivo model to study invasion and metastasis of human bladder carcinoma. Cancer Res. 41, 6660-6660 (1987).
- Heaney, J. A., Omellas, E. P., Daly, J. J., Lin, J. C., Rout, G. R. In vivo growth of human bladder cancer cell lines. Invest. Urol. 15, 380-380 (1978).
- Hadaschik, B., Black, P., Sea, J., Metwalli, A., Fazli, L., Dinney, C., Gleave, M., So, A. A validated mouse model for orthotopic bladder cancer using transurethral tumor inoculation and bioluminescence imaging. BJU International. 100, 1377-1384 (2007).
Tags
Mouse Bladder Wall InjectionOrthotopic StudyBladder PhenomenaStem CellSmooth MuscleCancer BiologySurgical Area CleaningAntiseptic SolutionSterilized Surgical EquipmentInhaled Isoflurane AnesthesiaBladder ExposureMidline Abdominal IncisionBladder DecompressionCell Suspension InjectionMicrosurgical Technique
Cite This Article
Fu, C., Apelo, C. A., Torres, B., Thai, K. H., Hsieh, M. H. Mouse Bladder Wall Injection. J. Vis. Exp. (53), e2523, doi:10.3791/2523 (2011).