Name: mouse model of surgically-induced endometriosis by auto-transplantation of uterine tissue
Uploaded: 2012-01-06T12:00:00
Description: Watch this Scientific Journal Video about Mouse Model of Surgically-induced Endometriosis by Auto-transplantation of Uterine Tissue at JoVE.com

Un agradecimiento especial a Chris Kassotis y Audrey Bailey para la revisión crítica de este manuscrito y al Dr. Scott Korte, Beeman Joseph, Curfman Alison, Kimball Pablo, Neibreggue Bridget, Redel Jacob, Schröder Amy, Steinberg Maija y Winkeler Stacey por su ayuda en la optimización de de este modelo en nuestro laboratorio. El financiamiento fue proporcionado por la capacitación clínica Biodetectives Grant (NIH T90) (KEP) de la Universidad de Missouri Ciencias de la Vida Programa de Licenciatura de Investigación de Oportunidades, MU Consejo de Investigación, Becas de investigación MU Junta y los NIH R21HD056441 (SCN).

1. La planificación de la cirugía de animales vivos <ol><li> Asegúrese de que la aprobación correspondiente se ha recibido para trabajar con animales de laboratorio. </li><li> Para los ratones y permitir que al menos una semana de aclimatación a su nuevo entorno. </li><li> Los ratones hembra que encuentra en la ausencia de exposición a las feromonas masculinas pueden dejar de pedalear, un fenómeno que se conoce como el efecto Whitten 15,16. A seguir adelante ratones transferencia empapados de orina m...

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E., Edwards, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+superovulation+and+pregnancy+in+mature+mice+by+gonadotrophins.">Induction of superovulation and pregnancy in mature mice by gonadotrophins.</a> J. Endocrinol. 15, 374-384 (1957).</li><li>Wilson, E. D., Zarrow, M. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+superovulation+in+the+immature+mouse+and+rat.">Comparison of superovulation in the immature mouse and rat.</a> J. Reprod. Fertil. 3, 148-158 (1962).</li><li>Lee, B., Du, H., Taylor, H. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+murine+endometriosis+induces+DNA+methylation+and+altered+gene+expression+in+eutopic+endometrium.">Experimental murine endometriosis induces DNA methylation and altered gene expression in eutopic endometrium.</a> Biol. Reprod. 80, 79-85 (2009).</li><li>Somigliana, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endometrial+ability+to+implant+in+ectopic+sites+can+be+prevented+by+interleukin-12+in+a+murine+model+of+endometriosis.">Endometrial ability to implant in ectopic sites can be prevented by interleukin-12 in a murine model of endometriosis.</a> Hum. Reprod. 14, 2944-2950 (1999).</li><li>Hirata, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+an+experimental+model+of+endometriosis+using+mice+that+ubiquitously+express+green+fluorescent+protein.">Development of an experimental model of endometriosis using mice that ubiquitously express green fluorescent protein.</a> Hum. Reprod. 20, 2092-2096 (2005).</li><li>Story, L., Kennedy, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+studies+in+endometriosis:+a+review.">Animal studies in endometriosis: a review.</a> Ilar. J. 45, 132-138 (2004).</li><li>Cummings, A. M., Hedge, J. M., Birnbaum, L. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+prenatal+exposure+to+TCDD+on+the+promotion+of+endometriotic+lesion+growth+by+TCDD+in+adult+female+rats+and+mice.">Effect of prenatal exposure to TCDD on the promotion of endometriotic lesion growth by TCDD in adult female rats and mice.</a> Toxicol. Sci. 52, 45-49 (1999).</li><li>Cummings, A. M., Metcalf, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+estrogen,+progesterone,+and+methoxychlor+on+surgically+induced+endometriosis+in+rats.">Effects of estrogen, progesterone, and methoxychlor on surgically induced endometriosis in rats.</a> Fundam. Appl. Toxicol. 27, 287-290 (1995).</li><li>Sharpe-Timms, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endometriotic+lesions+synthesize+and+secrete+a+haptoglobin-like+protein.">Endometriotic lesions synthesize and secrete a haptoglobin-like protein.</a> Biol. Reprod. 58, 988-994 (1998).</li><li>Yavuz, E., Oktem, M., Esinler, I., Toru, S. A., Zeyneloglu, H. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genistein+causes+regression+of+endometriotic+implants+in+the+rat+model.">Genistein causes regression of endometriotic implants in the rat model.</a> Fertil. Steril. 88, 1129-1134 (2007).</li><li>Dmitrieva, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endocannabinoid+involvement+in+endometriosis.">Endocannabinoid involvement in endometriosis.</a> Pain. 151, 703-710 (2010).</li><li>Efstathiou, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonsteroidal+antiinflammatory+drugs+differentially+suppress+endometriosis+in+a+murine+model.">Nonsteroidal antiinflammatory drugs differentially suppress endometriosis in a murine model.</a> Fertil. Steril. 83, 171-181 (2005).</li><li>Becker, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endostatin+inhibits+the+growth+of+endometriotic+lesions+but+does+not+affect+fertility.">Endostatin inhibits the growth of endometriotic lesions but does not affect fertility.</a> Fertil. Steril. 84, 1144-1155 (2005).</li><li>Becker, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Short+synthetic+endostatin+peptides+inhibit+endothelial+migration+in+vitro+and+endometriosis+in+a+mouse+model.">Short synthetic endostatin peptides inhibit endothelial migration in vitro and endometriosis in a mouse model.</a> Fertil. Steril. 85, 71-77 (2006).</li><li>Sharpe-Timms, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Using+rats+as+a+research+model+for+the+study+of+endometriosis.">Using rats as a research model for the study of endometriosis.</a> Ann. N.Y. Acad. Sci. 955, 318-327 (2002).</li><li>Stilley, J. A., Woods-Marshall, R., Sutovsky, M., Sutovsky, P., Sharpe-Timms, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduced+Fecundity+in+Female+Rats+with+Surgically+Induced+Endometriosis+and+in+Their+Daughters:+A+Potential+Role+for+Tissue+Inhibitors+of+Metalloproteinase+1.">Reduced Fecundity in Female Rats with Surgically Induced Endometriosis and in Their Daughters: A Potential Role for Tissue Inhibitors of Metalloproteinase 1.</a> Biol. Reprod. 80, (2009).</li></ol>

Hay varios parámetros críticos que deben tenerse en cuenta al realizar la inducción quirúrgica de la endometriosis en ratones. En primer lugar, la endometriosis es una enfermedad estrógeno dependiente y, como tal, esta cirugía se debe realizar en animales intactos o, alternativamente, en los animales suplementados con ovariectomizadas estrógenos 20. En segundo lugar, la sutura de la biopsia endometrial a la cascada arterial debe realizarse con extremo cuidado. Hemos encontrado que el uso de sólo dos n...

<table border="1"><tbody><tr><td> Nombre del reactivo / equipo </td><td> Empresa </td><td> Número de catálogo </td></tr><tr><td> Cera lápiz </td><td> Pescador </td><td> NC9954135 </td></tr><tr><td> Portaobjetos de vidrio </td><td> Pescador </td><td> 12-550-433 </td></tr><tr><td> Cuentagotas </td><td> Pescador </td><td> S79383 </td></tr><tr><td> Microscopio de luz estándar para la evaluación de frotis de citología vaginal </td><td></td><td></td></tr><tr><td> La buprenorfina HCL c3 (CarJet) 10X1ml </td><td> Animal mayordomo de la Salud de suministro </td><td> 022891 </td></tr><tr><td> Fosfato estéril (PBS) </td><td> Gibco </td><td> 14040-117 </td></tr><tr><td> 10.000 U / ml de penicilina, 10000 mg / ml de estreptomicina en el 0,85% de NaCl </td><td> Hyclone </td><td> SV30010 </td></tr><tr><td> Isoflurano </td><td> AbbottSanidad Animal </td><td> 05260-05 </td></tr><tr><td> El isoflurano no reinhalación anestesia sistema </td><td></td><td></td></tr><tr><td> Recirculación de agua caliente almohadilla eléctrica </td><td></td><td></td></tr><tr><td> 30 ml jeringa vaina </td><td> Pescador </td><td> 14 a 823-16G </td></tr><tr><td> Polvo estéril guantes sin </td><td> Fisherbrand </td><td> 19020558 </td></tr><tr><td> Oftálmicos pomada </td><td> Productos farmacéuticos importantes </td><td> 10033691 </td></tr><tr><td> Pequeñas tijeras eléctricas </td><td> Wahl </td><td> 9861-600 </td></tr><tr><td> Clorhexidina matorral </td><td> Pescador </td><td> NC9863042 </td></tr><tr><td> 70% de etanol </td><td></td><td></td></tr><tr><td> Campo estéril Polylined </td><td> Busse Desechables hospital </td><td> 696 </td></tr><tr><td> Tamaño 3 bisturí </td><td> Pescador </td><td> 22-079-657 </td></tr><tr><td> Número 10 hojas de bisturí </td><td> Pescador </td><td> 22-079-681 </td></tr><tr><td> Pequeñas tijeras quirúrgicas </td><td> Roboz </td><td> RS-5850 </td></tr><tr><td> Pequeña sierra semi-curva con fórceps </td><td> Roboz </td><td> RS-5135 </td></tr><tr><td> 5-0 negro trenzado de sutura de seda </td><td> Ethicon </td><td> K870H </td></tr><tr><td> Esterilizados cristal pyrex Petri </td><td> Corning </td><td> 70160-101 </td></tr><tr><td> 2 mm de biopsia por punción </td><td> Miltex </td><td> 33-31 </td></tr><tr><td> Una gasa estéril </td><td> Kendall </td><td> 1806 </td></tr><tr><td> 6-0 ETHILON * negro sutura monofilamento de nylon </td><td> Ethicon </td><td> 697G </td></tr><tr><td> Portaagujas (opcional) </td><td> Mundial de Instrumentos de Precisión </td><td> 500023 </td></tr><tr><td> 5-0 teñir trenzado recubierto vicryl sutura </td><td> Ethicon </td><td> J490G </td></tr><tr><td> 9mm autoclip herida clips </td><td> Becton Dickinson </td><td> 427631 </td></tr><tr><td> Autoclip aplicador y removedor </td><td> Becton Dickinson </td><td> 427630 </td></tr><tr><td> 23G aguja </td><td> Becton Dickinson </td><td> 305193 </td></tr><tr><td> 1cc jeringa </td><td> Becton Dickinson </td><td> 301025 </td></tr><tr><td> Aumento 5X soporte de vidrio (opcional) </td><td> Pescador </td><td> 14-648-23 </td></tr><tr><td> 10% de formol </td><td> Pescador </td><td> SF100-4 </td></tr><tr><td> Calibrador </td><td> Roboz </td><td> RS-6466 </td></tr><tr><td> Procesamiento / incrustación cassettes </td><td> Pescador </td><td> 15 a 197-700A </td></tr><tr><td> Biopsia de almohadillas de espuma </td><td> Pescador </td><td> 22-038-222 </td></tr><tr><td> RNAqueous ARN Kit de aislamiento </td><td>Ambion </td><td> AM1912 </td></tr><tr><td> Nitrógeno líquido </td><td></td><td></td></tr><tr><td> Ajustar tapa del tubo de microcentrífuga de la parte superior plana </td><td> Pescador </td><td> 02-681-240 </td></tr><tr><td> La ketamina (opcional) </td><td> Simga </td><td> K4138 </td></tr><tr><td> Domitor (medetomidina clorhidrato) (opcional) </td><td> Tocris </td><td> 2023 </td></tr><tr><td> Antisedan (atipamezol) (opcional) </td><td> Sigma </td><td> A9611 </td></tr></tbody></table>

mouse model of surgically-induced endometriosis by auto-transplantation of uterine tissue

La endometriosis es una enfermedad crónica y dolorosa, cuya etiología permanece desconocida. Además, el tratamiento de la endometriosis puede requerir la extirpación laparoscópica de lesiones y / o la gestión farmacéutica crónica del dolor y los síntomas de la infertilidad. El costo asociado con la endometriosis ha sido estimada en 22 mil millones de dólares por año en los Estados Unidos 1. Para avanzar en nuestra comprensión de los mecanismos subyacentes a esta enigmática enfermedad, los modelos animales han sido empleados. Los primates desarrollan espontáneamente endometriosis y por lo tanto, los modelos de primates más se asemejan a la enfermedad en las mujeres. Modelos de roedores, sin embargo, son más rentables y de fácil acceso 2. El modelo que se describe aquí implica una transferencia autóloga del tejido uterino en el mesenterio intestinal (Figura 1) y se desarrolló por primera vez en la rata 3 y luego a la del ratón 4. El objetivo del modelo de roedores autólogo de endometriosis quirúrgicamente inducido es para imitarla enfermedad en las mujeres. Nosotros y otros han demostrado previamente que el patrón de expresión de genes alterado observado en las lesiones de endometriosis de los ratones o las ratas espejos que se observa en las mujeres con la enfermedad 5,6. Una de las ventajas de realizar la cirugía en el ratón es que la abundancia de cepas de ratones transgénicos disponibles puede ayudar a los investigadores para determinar el papel de determinados componentes importantes en el establecimiento y crecimiento de la endometriosis. Un modelo alternativo en el que extirpar los fragmentos de endometrio humano se introducen en el peritoneo de los ratones inmunodeprimidos también es ampliamente utilizado, pero está limitada por la falta de un sistema inmune normal que se cree que es importante en la endometriosis 2,7. Es importante destacar que el modelo de ratón de la endometriosis inducida quirúrgicamente es un modelo versátil que se ha utilizado para estudiar cómo el sistema inmune 8, 9,10 hormonas y los factores ambientales afectan a 11,12 endometriosis, así como los efectos de la endometriosis en fertility 13 y el dolor 14.

Watch this Scientific Journal Video about Mouse Model of Surgically-induced Endometriosis by Auto-transplantation of Uterine Tissue at JoVE.com

Mouse Model of Surgically-induced Endometriosis by Auto-transplantation of Uterine Tissue

Una descripción de la inducción quirúrgica de la endometriosis en ratones y ratas por auto-trasplante de tejido uterino de la cascada arterial del mesenterio intestinal.

Modelo de ratón de la endometriosis quirúrgicamente inducida por la Auto-trasplante de tejido uterino

Endometriosis is a chronic, painful disease whose etiology remains unknown. Furthermore, treatment of endometriosis can require laparoscopic removal of ...

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Medicine

Mouse Model of Intraluminal MCAO: Cerebral Infarct Evaluation by Cresyl Violet Staining

Modelo de ratón de MCAO intraluminal: Evaluación del infarto cerebral mediante tinción con violeta de cresilo

Stroke is the third  cause of mortality and the leading cause of disability in the World. Ischemic  stroke accounts for approximately 80% of all strokes. ...

Investigación

Medicina

Implantation of Inferior Vena Cava Interposition Graft in Mouse Model

Implantación de la vena cava inferior interposición del injerto en modelo de ratón

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. ...

Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation

El trasplante de la válvula pulmonar Utilizando un modelo de ratón de heterotópico trasplante cardiaco

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed ...

Rose Bengal Photothrombosis by Confocal Optical Imaging In Vivo: A Model of Single Vessel Stroke

Rose Bengal Photothrombosis by Confocal Optical Imaging In Vivo:  A Model of Single Vessel Stroke

Rosa de Bengala Photothrombosis por imagen óptica confocal<em&gt; En Vivo</em&gt;: Un modelo de solo recipiente Stroke

In vivo imaging techniques have increased in utilization due to recent advances in imaging dyes and optical technologies, allowing for the ability to ...

Assessment of Cardiac Morphological and Functional Changes in Mouse Model of Transverse Aortic Constriction by Echocardiographic Imaging

Evaluación de la cardíaco morfológica y funcional Los cambios en el modelo de ratón de la constricción de la aorta transversa por ecocardiográfico Imaging

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable ...

Creation of a Rodent Model of Abdominal Aortic Aneurysm by Blocking Adventitial Vasa Vasorum Perfusion

Creación de un modelo de roedor del Aneurysm aórtico Abdominal bloqueando Adventitial de la Vasa Vasorum de la perfusión

The adventitial vasa vasorum (VV) provides oxygen and nourishment to the aortic wall. Hypoxia in the aortic wall can cause enlarged abdominal aortic ...

Contractility Measurements of Human Uterine Smooth Muscle to Aid Drug Development

Mediciones de la contractilidad del musculo liso uterino humano desarrollo de fármacos de ayuda

Discovery and characterization of novel pharmaceutical compounds or biochemical probes rely on robust and physiologically relevant assay systems. We ...

A Mouse Model of Orthopedic Surgery to Study Postoperative Cognitive Dysfunction and Tissue Regeneration

Un modelo de ratón de cirugía ortopédica para el estudio de la disfunción cognitiva postoperatoria y regeneración de los tejidos

Surgery is commonly used to improve and maintain quality of life. Unfortunately, in vulnerable patients such as the elderly, complications may occur and ...

Improvement of a Closed Chest Porcine Myocardial Infarction Model by Standardization of Tissue and Blood Sampling Procedures

Mejora de un modelo de infarto de miocardio porcino pecho cerrado por normalización del tejido y procedimientos de muestreo de sangre

Myocardial ischemia reperfusion (I/R) injury contributes to almost half of the necrotic area after myocardial infarction. To date there is no approved ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

Medición de la contractilidad isométrica de la arteria mesentérica de ratón con cable miografía

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...