Un ringraziamento speciale a Chris Kassotis e Audrey Bailey per la revisione critica di questo manoscritto e il Dr. Scott Korte, Giuseppe Beeman, Alison Curfman, Paul Kimball, Bridget Neibreggue, Jacob Redel, Amy Schroder, Maija Steinberg, e Stacey Winkeler per la loro assistenza per l&#39;ottimizzazione di questo modello nel nostro laboratorio. Il finanziamento è stato fornito dalla formazione di Grant Clinica Biodetectives (NIH T90) (Kep), Università del Missouri Scienze della Vita Undergraduate Research Program Opportunità, MU Research Council, MU Assegni di ricerca Board e NIH R21HD056441 (SCN).

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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. 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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. 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Nessun conflitto di interessi dichiarati.

Ci sono diversi parametri critici che dovrebbero essere notato durante l&#39;esecuzione l&#39;induzione chirurgica di endometriosi nei topi. In primo luogo, l&#39;endometriosi è una malattia estrogeno-dipendenti e come tale questo tipo di chirurgia deve essere eseguita nell&#39;animale intatto o in alternativa negli animali ovariectomizzati integrato con estrogeni 20. In secondo luogo, la sutura biopsie endometriali alla cascata arteriosa deve essere eseguita con estrema cura. Abbiamo scoperto che usando sol...

<table border="1"><tbody><tr><td> Nome del reagente / apparecchiatura </td><td> Azienda </td><td> Numero di catalogo </td></tr><tr><td> Matita di cera </td><td> Pescatore </td><td> NC9954135 </td></tr><tr><td> Vetrini </td><td> Pescatore </td><td> 12-550-433 </td></tr><tr><td> Contagocce </td><td> Pescatore </td><td> S79383 </td></tr><tr><td> Microscopio ottico standard per la valutazione strisci citologia vaginale </td><td></td><td></td></tr><tr><td> Buprenorfina HCL c3 (CARJET) 10X1ml </td><td> Butler animali Alimentazione Salute </td><td> 022891 </td></tr><tr><td> Fosfato sterile salina tamponata (PBS) </td><td> Gibco </td><td> 14040-117 </td></tr><tr><td> 10.000 U / ml penicillina, 10.000 mg / ml di streptomicina in 0,85% NaCl </td><td> Hyclone </td><td> SV30010 </td></tr><tr><td> Isoflurano </td><td> AbbottAnimal Health </td><td> 05260-05 </td></tr><tr><td> Isoflurano non rirespirazione anestetico sistema </td><td></td><td></td></tr><tr><td> Ricircolo di acqua calda di riscaldamento pad </td><td></td><td></td></tr><tr><td> 30 ml siringa guaina </td><td> Pescatore </td><td> 14-823-16G </td></tr><tr><td> Polvere di guanti sterili libero </td><td> Fisherbrand </td><td> 19020558 </td></tr><tr><td> Unguento oftalmico </td><td> Farmaceutica principali </td><td> 10033691 </td></tr><tr><td> Piccolo tagliaunghie elettrici </td><td> Wahl </td><td> 9861-600 </td></tr><tr><td> Clorexidina macchia </td><td> Pescatore </td><td> NC9863042 </td></tr><tr><td> 70% di etanolo </td><td></td><td></td></tr><tr><td> Polylined campo sterile </td><td> Busse Ospedale Disposables </td><td> 696 </td></tr><tr><td> Taglia 3 bisturi </td><td> Pescatore </td><td> 22-079-657 </td></tr><tr><td> Numero 10 lame bisturi </td><td> Pescatore </td><td> 22-079-681 </td></tr><tr><td> Piccole forbici chirurgiche </td><td> Roboz </td><td> RS-5850 </td></tr><tr><td> Piccoli semi-curve seghettate forcipe </td><td> Roboz </td><td> RS-5135 </td></tr><tr><td> 5-0 di seta nera intrecciata sutura </td><td> Ethicon </td><td> K870H </td></tr><tr><td> Sterilizzato vetro pirex Petri </td><td> Corning </td><td> 70160-101 </td></tr><tr><td> 2 millimetri biopsia </td><td> Miltex </td><td> 33-31 </td></tr><tr><td> Garza sterile </td><td> Kendall </td><td> 1806 </td></tr><tr><td> 6-0 nero ethilon sutura monofilo di nylon </td><td> Ethicon </td><td> 697G </td></tr><tr><td> Driver ago (opzionale) </td><td> Mondo Strumenti di precisione </td><td> 500023 </td></tr><tr><td> 5-0 undyed intrecciato rivestito vicryl sutura </td><td> Ethicon </td><td> J490G </td></tr><tr><td> 9 millimetri Autoclip ferita clip </td><td> Becton Dickinson </td><td> 427631 </td></tr><tr><td> Autoclip applicatore e di rimozione </td><td> Becton Dickinson </td><td> 427630 </td></tr><tr><td> 23G ago </td><td> Becton Dickinson </td><td> 305193 </td></tr><tr><td> 1cc siringa </td><td> Becton Dickson </td><td> 301025 </td></tr><tr><td> 5X vetro stare ingrandimento (opzionale) </td><td> Pescatore </td><td> 14-648-23 </td></tr><tr><td> 10% di formalina tamponata </td><td> Pescatore </td><td> SF100-4 </td></tr><tr><td> Pinze </td><td> Roboz </td><td> RS-6466 </td></tr><tr><td> Elaborazione / embedding cassette </td><td> Pescatore </td><td> 15-197-700A </td></tr><tr><td> Biopsia cuscinetti in schiuma </td><td> Pescatore </td><td> 22-038-222 </td></tr><tr><td> RNAqueous RNA Kit di isolamento </td><td>Ambion </td><td> AM1912 </td></tr><tr><td> Azoto liquido </td><td></td><td></td></tr><tr><td> Snap microcentrifuga coppola tubo orizzontale </td><td> Pescatore </td><td> 02-681-240 </td></tr><tr><td> Ketamina (opzionale) </td><td> Simga </td><td> K4138 </td></tr><tr><td> Domitor (medetomidina cloridrato) (opzionale) </td><td> Tocris </td><td> 2023 </td></tr><tr><td> Antisedan (atipamezolo) (opzionale) </td><td> Sigma </td><td> A9611 </td></tr></tbody></table>

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

L&#39;endometriosi è una malattia cronica, dolorosa malattia la cui eziologia rimane sconosciuta. Inoltre, il trattamento di endometriosi può richiedere la rimozione laparoscopica delle lesioni, e / o croniche gestione farmaceutica del dolore e dei sintomi infertilità. Il costo associato con endometriosi è stato stimato a 22 miliardi di dollari all&#39;anno negli Stati Uniti 1. Per ulteriormente la nostra comprensione dei meccanismi alla base di questa malattia enigmatica, i modelli animali sono stati impiegati. Primati sviluppano spontaneamente endometriosi e quindi modelli di primati più simili alla malattia nelle donne. Modelli di roditori, tuttavia, sono più convenienti e facilmente reperibili 2. Il modello che abbiamo descritto qui comporta un trasferimento autologo di tessuto uterino al mesentere intestinale (Figura 1) ed è stato sviluppato nel ratto 3 e poi trasferito al mouse 4. L&#39;obiettivo del modello roditore autologo di endometriosi chirurgicamente indotta è quella di imitarela malattia nelle donne. Noi e altri hanno già dimostrato che l&#39;alterato pattern di espressione genica osservata nelle lesioni endometriosiche da topi o ratti rispecchia quella osservata nelle donne con la malattia 5,6. Uno dei vantaggi del realizzare l&#39;intervento nel topo è che l&#39;abbondanza di ceppi di topi transgenici disponibile può aiutare i ricercatori a determinare il ruolo di specifici componenti importanti nella creazione e la crescita di endometriosi. Un modello alternativo in cui vengono introdotti asportati frammenti umani endometriale al peritoneo di topi immunocompromessi è anche ampiamente utilizzato, ma è limitata dalla mancanza di un normale sistema immunitario che si ritiene essere importante per l&#39;endometriosi 2,7. È importante sottolineare che il modello murino di endometriosi indotta chirurgicamente è un modello versatile che è stato utilizzato per studiare come il sistema immunitario 8, 9,10 ormoni e fattori ambientali influenzano 11,12 endometriosi così come gli effetti dell&#39;endometriosi sulla Fertility 13 e il dolore 14.

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

Una descrizione della induzione chirurgica di endometriosi in topi e ratti con auto-trapianto di tessuto uterino alla cascata delle arterie del mesentere intestinale.

Modello murino di endometriosi chirurgicamente indotto da Auto-trapianto di tessuto uterino

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

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28.5K Views.  University of Missouri. Una descrizione della induzione chirurgica di endometriosi in topi e ratti con auto-trapianto di tessuto uterino alla cascata delle arterie del mesentere intestinale.

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

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

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. However, the thrombolytic tissue plasminogen activator (tPA) is the only treatment of acute ischemic stroke that exists. This led researchers to develop several ischemic stroke models in a variety of species. Two major types of rodent models have been developed: models of global cerebral ischemia or focal cerebral ischemia. To mimic ischemic stroke in patients, in whom approximately 80% thrombotic or embolic strokes occur in the territory of the middle cerebral artery (MCA), the intraluminal middle cerebral artery occlusion (MCAO) model is quite relevant for stroke studies. This model was first developed in rats by Koizumi et al. in 1986 1. Because of the ease of genetic manipulation in mice, these models have also been developed in this species 2-3.
	Herein, we present the transient MCA occlusion procedure in C57/Bl6 mice. Previous studies have reported that physical properties of the occluder such as tip diameter, length, shape, and flexibility are critical for the reproducibility of the infarct volume 4. Herein, a commercial silicon coated monofilaments (Doccol Corporation) have been used. Another great advantage is that this monofilament reduces the risk to induce subarachnoid hemorrhages. Using the Zeiss stereo-microscope Stemi 2000, the silicon coated monofilament was introduced into the internal carotid artery (ICA) via a cut in the external carotid artery (ECA) until the monofilament occludes the base of the MCA. Blood flow was restored 1 hour later by removal of the monofilament to mimic the restoration of blood flow after lysis of a thromboembolic clot in humans. The extent of cerebral infarct may be evaluated first by a neurologic score and by the measurement of the infarct volume. Ischemic mice were thus analyzed for their neurologic score at different post-reperfusion times. To evaluate the infarct volume, staining with 2,3,5-triphenyltetrazolium chloride (TTC) was usually performed. Herein, we used cresyl violet staining since it offers the opportunity to test many critical markers by immunohistochemistry. In this video, we report the MCAO procedure; neurological scores and the evaluation of the infarct volume by cresyl violet staining.

The intraluminal middle cerebral occlusion model in mice is herein presented. The extent of cerebral infarct is evaluated by a neurologic score and cresyl violet staining, an alternative staining to TTC, offering the great advantage to test in parallel many interest markers.

Modello murino di MCAO intraluminale: valutazione infarto cerebrale da colorazione viola cresile

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. ...

Ricerca

Medicina

Implantation of Inferior Vena Cava Interposition Graft in Mouse Model

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. The long-term clinical results showed excellent patency rates, however, with significant incidence of stenosis. To investigate the cellular and molecular mechanisms of vascular neotissue formation and prevent stenosis development in tissue engineered vascular grafts (TEVGs), we developed a mouse model of the graft with approximately 1 mm internal diameter. First, the TEVGs were assembled from biodegradable tubular scaffolds fabricated from a polyglycolic acid nonwoven felt mesh coated with &#949;-caprolactone and L-lactide copolymer. The scaffolds were then placed in a lyophilizer, vacuumed for 24 hr, and stored in a desiccator until cell seeding. Second, bone marrow was collected from donor mice and mononuclear cells were isolated by density gradient centrifugation. Third, approximately one million cells were seeded on a scaffold and incubated O/N. Finally, the seeded scaffolds were then implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. The implanted grafts demonstrated excellent patency (&#62;90%) without evidence of thromboembolic complications or aneurysmal formation. This murine model will aid us in understanding and quantifying the cellular and molecular mechanisms of neotissue formation in the TEVG.

To improve our knowledge of cellular and molecular neotissue formation, a murine model of the TEVG was recently developed. The grafts were implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. This model achieves similar results to those achieved in our clinical investigation, but over a far shortened time-course.

L&#39;impianto di Vena cava inferiore Interposizione Graft in modello murino

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

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed results. However, the cellular and molecular mechanisms of the neotissue development, valve thickening, and stenosis development are not researched extensively. To answer the above questions, we developed a murine heterotopic heart valve transplantation model. A heart valve was harvested from a valve donor mouse and transplanted to a heart donor mouse. The heart with a new valve was transplanted heterotopically to a recipient mouse. The transplanted heart showed its own heartbeat, independent of the recipient&#8217;s heartbeat. The blood flow was quantified using a high frequency ultrasound system with a pulsed wave Doppler. The flow through the implanted pulmonary valve showed forward flow with minimal regurgitation and the peak flow was close to 100 mm/sec. This murine model of heart valve transplantation is highly versatile, so it can be modified and adapted to provide different hemodynamic environments and/or can be used with various transgenic mice to study neotissue development in a tissue engineered heart valve.

In order to understand the cellular and molecular mechanisms underlying neotissue formation and stenosis development in tissue engineered heart valves, a murine model of heterotopic heart valve transplantation was developed. A pulmonary heart valve was transplanted to recipient using the heterotopic heart transplantation technique.

Il trapianto di valvola polmonare Utilizzando un modello murino di trapianto cardiaco eterotopico

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

Utero-tubal Embryo Transfer and Vasectomy in the Mouse Model

The transfer of preimplantation embryos to a surrogate female is a required step for the production of genetically modified mice or to study the effects of epigenetic alterations originated during preimplantation development on subsequent fetal development and adult health. The use of an effective and consistent embryo transfer technique is crucial to enhance the generation of genetically modified animals and to determine the effect of different treatments on implantation rates and survival to term. Embryos at the blastocyst stage are usually transferred by uterine transfer, performing a puncture in the uterine wall to introduce the embryo manipulation pipette. The orifice performed in the uterus does not close after the pipette has been withdrawn, and the embryos can outflow to the abdominal cavity due to the positive pressure of the uterus. The puncture can also produce a hemorrhage that impairs implantation, blocks the transfer pipette and may affect embryo development, especially when embryos without zona are transferred. Consequently, this technique often results in very variable and overall low embryo survival rates. Avoiding these negative effects, utero-tubal embryo transfer take advantage of the utero-tubal junction as a natural barrier that impedes embryo outflow and avoid the puncture of the uterine wall. Vasectomized males are required for obtaining pseudopregnant recipients. A technique to perform vasectomy is described as a complement to the utero-tubal embryo transfer.

Utero-tubal embryo transfer uses the utero-tubal junction as a barrier to prevent the embryo outflow that may occur when performing uterine transfer. Vasectomized males are required to obtain pseudopregnant recipients for embryo transfer. Both techniques are discussed.

Trasferimento di Embrione utero-tubarica e vasectomia nel modello murino

The transfer of preimplantation embryos to a surrogate female is a required step for the production of genetically modified mice or to study the effects ...

Biologia

A Syngeneic Murine Model of Endometriosis using Naturally Cycling Mice

Endometriosis is a leading cause of pelvic pain and infertility. It is defined by the presence of endometrial tissue in extrauterine locations. The development of novel therapies and diagnostic tools for endometriosis has been limited due in part to challenges in studying the disease. Outside of primates, few mammals menstruate, and none develop spontaneous endometriosis. Rodent models are popular but require artificial induction of endometriosis, with many utilizing either immunocompromised mice or surgically induced disease. Recently, more attention has been given to models involving intraperitoneal injection. We present a murine model of endometriosis that integrates several features of existing endometriosis models into a novel, simplified system that relies on microscopic quantification in lieu of subjective grading. In this model, we perform hormonal stimulation of donor mice, intraperitoneal injection, systematic abdominal survey and tissue harvest, and histologic quantification that can be performed and verified at any time after necropsy. This model requires minimal resources and training; does not require expertise by lab technicians in murine survival surgery or in the identification of gross endometriotic lesions; can be used in immunocompromised, immunocompetent, and/or mutant mice; and reliably creates endometriotic lesions that are histologically consistent with human endometriotic disease.

Many rodent models of endometriosis are limited by technical complexity, reproducibility, and/or need for immunocompromised animals or special reporter mice. We present a simplified system of lesion induction using any experimental mouse with an independently verifiable, objective scoring system and with no requirement for ovariectomy or survival surgery.

Un modello murino singenico di endometriosi utilizzando topi naturalmente ciclabili

Endometriosis is a leading cause of pelvic pain and infertility. It is defined by the presence of endometrial tissue in extrauterine locations. The ...

Noninvasive Monitoring of Lesion Size in a Heterologous Mouse Model of Endometriosis

Here, we describe a protocol for the implementation of a heterologous mouse model in which progression of endometriosis can be assessed in real time through noninvasive monitoring of fluorescence emitted by implanted ectopic human endometrial tissue. For this purpose, biopsies of human endometrium are obtained from donor women ongoing oocyte donation. Human endometrial fragments are cultured in the presence of adenoviruses engineered to express cDNA for the reporter fluorescent protein mCherry. Upon visualization, labeled tissues with an optimal rate of fluorescence after infection are subsequently chosen for the implantation in recipient mice. One week prior to the implantation surgery, recipient mice are oophorectomized, and estradiol pellets are placed subcutaneously to sustain the survival and growth of lesions. On the day of surgery mice are anesthetized, and peritoneal cavity accessed through a small (1.5 cm) incision by the linea-alba. Fluorescently labeled implants are tweezed, briefly soaked in glue and attached to the peritoneal layer. Incisions are sutured, and animals left to recover for a couple of days. Fluorescence emitted by endometriotic implants is usually non-invasively monitored every 3 days for 4 weeks with an in vivo imaging system. Variations in the size of endometriotic implants can be estimated in real time by quantification of the mCherry signal and normalization against the initial time-point showing maximal fluorescence intensity.
Traditional preclinical rodents of models of endometriosis do not allow non-invasive monitoring of lesion in real time but rather allow evaluation of the effects of drugs assayed at the end point. This protocol allows one to track lesions in real time and is more useful to explore the therapeutic potential of drugs in preclinical models of endometriosis. The main limitation of the model thus generated is that non-invasive monitoring is not possible over long periods of time due to the episomal expression of Ad-virus.

Here, we present a protocol for live-imaging of fluorescently labeled human endometrial fragments grafted in mice. The method allows studying the effects of drugs of choice on endometriotic lesion size through monitoring and quantification of fluorescence emitted by the fluorescent reporter on real time

Monitoraggio non invasivo della dimensione della lesione in un modello murino eterologo di endometriosi

Here, we describe a protocol for the implementation of a heterologous mouse model in which progression of endometriosis can be assessed in real time ...

Development of a Novel Endometrioma-Simulating Mouse Model

Establishment of an Experimental Mouse Model of Endometrioma to Study its Related Infertility

Endometrioma (OMA), a subtype of endometriosis characterized by the formation of endometriotic cysts in the ovaries, affects 17-44% of individuals diagnosed with endometriosis. Women with OMA often experience compromised fertility, yet the exact mechanisms underlying OMA-associated infertility remain unclear. Notably, existing animal models simulate superficial peritoneal endometriosis (SUP) and deep infiltrating endometriosis (DIE), leaving a notable gap in research focused on OMA. In response to the gap of knowledge, this paper introduces a pioneering OMA-simulating mouse model and provides a comprehensive description of the techniques and procedures employed in the model. With a high success rate of 83% and ovarian lesion specificity, this model holds significant promise for advancing our understanding of OMA, particularly in the context of infertility. It offers a valuable platform for conducting targeted research into OMA-associated fertility challenges, potentially paving the way for improved diagnostic and therapeutic strategies in the field of reproductive medicine.

This study introduces a novel mouse model specifically designed to simulate endometrioma, a clinically significant subtype of endometriosis. By utilizing C57BL/6J mice, the model aims to elucidate the pathophysiological mechanisms underlying endometrioma-related infertility, offering a refined tool to bridge the current knowledge gap in reproductive medicine.

Endometrioma (OMA), a subtype of endometriosis characterized by the formation of endometriotic cysts in the ovaries, affects 17-44% of individuals ...

Establishing 3D Endometrial Organoids from the Mouse Uterus

Endometrial tissue lines the inner cavity of the uterus and is under the cyclical control of estrogen and progesterone. It is a tissue that is composed of luminal and glandular epithelium, a stromal compartment, a vascular network, and a complex immune cell population. Mouse models have been a powerful tool to study the endometrium, revealing critical mechanisms that control implantation, placentation, and cancer. The recent development of 3D endometrial organoid cultures presents a state-of-the-art model to dissect the signaling pathways that underlie endometrial biology. Establishing endometrial organoids from genetically engineered mouse models, analyzing their transcriptomes, and visualizing their morphology at a single-cell resolution are crucial tools for the study of endometrial diseases. This paper outlines methods to establish 3D cultures of endometrial epithelium from mice and describes techniques to quantify gene expression and analyze the histology of the organoids. The goal is to provide a resource that can be used to establish, culture, and study the gene expression and morphological characteristics of endometrial epithelial organoids.

This protocol describes methodologies to establish mouse endometrial epithelial organoids for gene expression and histological analyses.

Stabilire organoidi endometriali 3D dall'utero del topo

Endometrial tissue lines the inner cavity of the uterus and is under the cyclical control of estrogen and progesterone. It is a tissue that is composed of ...

Optimized Thin Endometrium Mouse Model with Low Mortality and Minimal Adhesions

Establishing a Mouse Model of Thin Endometrium

Thin endometrium (TE) has been widely recognized as a critical cause of infertility. However, the pathogenesis of TE remains unclear, and satisfactory treatment options are still urgently needed. Several animal models of TE have been developed, but the mouse model involving abdominal surgery and injection of 95% ethanol presents a formidable challenge due to the high mortality rate and risk of intrauterine adhesions if not performed correctly. Here, we describe a detailed protocol that produces reliable and reproducible TE with a very low mortality rate and minimal intrauterine adhesions by injecting 95% ethanol into the mouse uterus with varying infusion times. The results showed that all of the mice successfully developed TE with infusion times ranging from 1-3 min, characterized by a typical reduction in endometrial thickness and the number of glands, as well as excessive endometrial fibrosis. These findings suggest that this mouse model is suitable for studying thin endometrium and can serve as a platform for developing future TE treatments.

This article describes a detailed protocol for producing a reliable and reproducible thin endometrium with a very low mortality rate and minimal intrauterine adhesions by injecting 95% ethanol into the mouse uterus within 1-3 min.

Thin endometrium (TE) has been widely recognized as a critical cause of infertility. However, the pathogenesis of TE remains unclear, and satisfactory ...

Modeling Metastatic Endometrial Cancer  Model: A Technique to Generate Endometrial Cancer with Lymph Node Metastasis in Rabbit Models

Use a number 11 scalpel blade to make a 2.5-centimeter incision, 1 centimeter cranial to the symphysis pubis through skin and subcutaneous tissue. Incise the rectus fascia and dissect the rectus muscles laterally to expose the underlying peritoneum. Confirm that the under surface is clear of bowel or other abdominal organs. Sharply enter the peritoneum to identify the urinary bladder and sweep a gloved finger superiorly, posteriorly, and laterally over the apex of the bladder to locate the uterine horns.
Extract the uterine horns through the abdominal incision to rest on the abdominal wall. Use a 3-0 braided absorbable suture to perform a single suture ligation of each uterine horn, approximately 1.5 to 2 centimeters distal to the cervices and just medial to the uterine arteries. Then, tie the sutures snugly to occlude the distal uterine horns.
To inoculate the myometrium with the previously prepared VX2 cell suspension, load a 1-milliliter syringe equipped with a 27 gauge needle with 1 milliliter of cells. Slowly inject 0.5 milliliters of the cell suspension over 1 minute into each uterine horn proximal to the suture site between the suture and the cervix. Then, apply pressure to the myometrial injection site for 30 seconds to minimize cell leakage. Inspect the injection and suture sites for hemostasis before placing the uterine horns back into the abdomen.

Modellazione del modello di cancro dell'endometrio metastatico: una tecnica per generare il cancro dell'endometrio con metastasi linfonodali in modelli di coniglio