Name: Mouse Model of Surgically-induced Endometriosis by Auto-transplantation of Uterine Tissue (Video) | JoVE Journal
Uploaded: 2012-01-06T12:00:00.000+00:00
Duration: 8 min 2 s
Description: Learn to surgically induce endometriosis in mice by auto-transplanting uterine tissue to the intestinal mesentery. This model creates endometrioid lesions resembling human disease for studying pathophysiology and therapeutics.

ai_agent_IP

ai_agent

maintenance

instantTrialDurationMinutes

isBiopharma

enableNewMyJoveSectionDesign

educationFeatures

seoMetaApi

ai_copilot_institution_blacklist

ai_chat_institution_blacklist

freeAccessWindow

ssoRoleCapture

scheduledMaintenanceBanner

joveLabsFeatures

disable_autoplay_for_bots

extended_trial_duration

5 minutes

Special thanks to Chris Kassotis and Audrey Bailey for critical review of this manuscript and to Dr. Scott Korte, Joseph Beeman, Alison Curfman, Paul Kimball, Bridget Neibreggue, Jacob Redel, Amy Schroder, Maija Steinberg, and Stacey Winkeler for their assistance in optimization of this model in our laboratory. Funding was provided by the Clinical Biodetectives Training Grant (NIH T90) (KEP), University of Missouri Life Sciences Undergraduate Research Opportunities Program, MU Research Council, MU Research Board grants and NIH R21HD056441 (SCN).

<ol><li>Simoens, S., Hummelshoj, L., D'Hooghe, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Endometriosis%3A+cost+estimates+and+methodological+perspective%5BTitle%5D)+AND+%22Hum.+Reprod.+Update%22%5BJournal%5D)">Endometriosis: cost estimates and methodological perspective.</a> Hum. Reprod. Update. 13, 395-404 (2007).</li>
<li>Grummer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Animal+models+in+endometriosis+research%5BTitle%5D)+AND+%22Hum.+Reprod.+Update%22%5BJournal%5D)">Animal models in endometriosis research.</a> Hum. Reprod. Update. 12, 641-649 (2006).</li>
<li>Vernon, M. W., Wilson, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Studies+on+the+surgical+induction+of+endometriosis+in+the+rat%5BTitle%5D)+AND+%22Fertil.+Steril%22%5BJournal%5D)">Studies on the surgical induction of endometriosis in the rat.</a> Fertil. Steril. 44, 684-694 (1985).</li>
<li>Cummings, A. M., Metcalf, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Induction+of+endometriosis+in+mice%3A+a+new+model+sensitive+to+estrogen%5BTitle%5D)+AND+%22Reprod.+Toxicol%22%5BJournal%5D)">Induction of endometriosis in mice: a new model sensitive to estrogen.</a> Reprod. Toxicol. 9, 233-238 (1995).</li>
<li>Pelch, K. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Aberrant+gene+expression+profile+in+a+mouse+model+of+endometriosis+mirrors+that+observed+in+women%5BTitle%5D)+AND+%22Fertil.+Steril%22%5BJournal%5D)">Aberrant gene expression profile in a mouse model of endometriosis mirrors that observed in women.</a> Fertil. Steril. 93, 1615-1627 (2010).</li>
<li>Flores, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Molecular+profiling+of+experimental+endometriosis+identified+gene+expression+patterns+in+common+with+human+disease%5BTitle%5D)+AND+%22Fertil.+Steril%22%5BJournal%5D)">Molecular profiling of experimental endometriosis identified gene expression patterns in common with human disease.</a> Fertil. Steril. 87, 1180-1199 (2007).</li>
<li>Giudice, L. C., Kao, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Endometriosis%5BTitle%5D)+AND+%22Lancet%22%5BJournal%5D)">Endometriosis.</a> Lancet. 364, 1789-1799 (2004).</li>
<li>Lin, Y. J., Lai, L. ei, Y, H., Wing, L. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Neutrophils+and+macrophages+promote+angiogenesis+in+the+early+stage+of+endometriosis+in+a+mouse+model%5BTitle%5D)+AND+%22Endocrinology%22%5BJournal%5D)">Neutrophils and macrophages promote angiogenesis in the early stage of endometriosis in a mouse model.</a> Endocrinology. 147, 1278-1286 (2006).</li>
<li>Fang, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Intact+progesterone+receptors+are+essential+to+counteract+the+proliferative+effect+of+estradiol+in+a+genetically+engineered+mouse+model+of+endometriosis%5BTitle%5D)+AND+%22Fertil.+Steril%22%5BJournal%5D)">Intact progesterone receptors are essential to counteract the proliferative effect of estradiol in a genetically engineered mouse model of endometriosis.</a> Fertil. Steril. 82, 673-678 (2004).</li>
<li>Fang, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Genetic+or+enzymatic+disruption+of+aromatase+inhibits+the+growth+of+ectopic+uterine+tissue%5BTitle%5D)+AND+%22J.+Clin.+Endocrinol.+Metab%22%5BJournal%5D)">Genetic or enzymatic disruption of aromatase inhibits the growth of ectopic uterine tissue.</a> J. Clin. Endocrinol. Metab. 87, 3460-3466 (2002).</li>
<li>Cummings, A. M., Metcalf, J. L., Birnbaum, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Promotion+of+endometriosis+by+2%2C3%2C7%2C8-tetrachlorodibenzo-p-dioxin+in+rats+and+mice%3A+time-dose+dependence+and+species+comparison%5BTitle%5D)+AND+%22Toxicol.+Appl.+Pharmacol%22%5BJournal%5D)">Promotion of endometriosis by 2,3,7,8-tetrachlorodibenzo-p-dioxin in rats and mice: time-dose dependence and species comparison.</a> Toxicol. Appl. Pharmacol. 138, 131-139 (1996).</li>
<li>Foster, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Morphologic+characteristics+of+endometriosis+in+the+mouse+model%3A+application+to+toxicology%5BTitle%5D)+AND+%22Can.+J.+Physiol.+Pharmacol%22%5BJournal%5D)">Morphologic characteristics of endometriosis in the mouse model: application to toxicology.</a> Can. J. Physiol. Pharmacol. 75, 1188-1196 (1997).</li>
<li>Cummings, A. M., Metcalf, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effect+of+surgically+induced+endometriosis+on+pregnancy+and+effect+of+pregnancy+and+lactation+on+endometriosis+in+mice%5BTitle%5D)+AND+%22Proc.+Soc.+Exp.+Biol.+Med%22%5BJournal%5D)">Effect of surgically induced endometriosis on pregnancy and effect of pregnancy and lactation on endometriosis in mice.</a> Proc. Soc. Exp. Biol. Med. 212, 332-337 (1996).</li>
<li>Lu, Y., Nie, J., Liu, X., Zheng, Y., Guo, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Trichostatin+A%2C+a+histone+deacetylase+inhibitor%2C+reduces+lesion+growth+and+hyperalgesia+in+experimentally+induced+endometriosis+in+mice%5BTitle%5D)+AND+%22Hum.+Reprod%22%5BJournal%5D)">Trichostatin A, a histone deacetylase inhibitor, reduces lesion growth and hyperalgesia in experimentally induced endometriosis in mice.</a> Hum. Reprod. 25, 1014-1025 (2010).</li>
<li>Whitten, W. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Modification+of+the+oestrous+cycle+of+the+mouse+by+external+stimuli+associated+with+the+male%5BTitle%5D)+AND+%22J.+Endocrinol%22%5BJournal%5D)">Modification of the oestrous cycle of the mouse by external stimuli associated with the male.</a> J. Endocrinol. 13, 399-404 (1956).</li>
<li>Whitten, W. K., Bronson, F. H., Greenstein, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Estrus-inducing+pheromone+of+male+mice%3A+transport+by+movement+of+air%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">Estrus-inducing pheromone of male mice: transport by movement of air.</a> Science. 161, 584-585 (1968).</li>
<li>Goldman, J. M., Murr, A. S., Cooper, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+rodent+estrous+cycle%3A+characterization+of+vaginal+cytology+and+its+utility+in+toxicological+studies%5BTitle%5D)+AND+%22Birth.+Defects.+Res.+B.+Dev.+Reprod.+Toxicol%22%5BJournal%5D)">The rodent estrous cycle: characterization of vaginal cytology and its utility in toxicological studies.</a> Birth. Defects. Res. B. Dev. Reprod. Toxicol. 80, 84-97 (2007).</li>
<li>Pritchett-Corning, K. R., Mulder, G. B., Luo, Y., White, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Principles+of+Rodent+Surgery+for+the+New+Surgeon%5BTitle%5D)+AND+%22J.+Vis.+Exp%22%5BJournal%5D)">Principles of Rodent Surgery for the New Surgeon.</a> J. Vis. Exp. (47), e2586-e2586 (2011).</li>
<li>Moen, M. H., Halvorsen, T. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Histologic+confirmation+of+endometriosis+in+different+peritoneal+lesions%5BTitle%5D)+AND+%22Acta.+Obstet.+Gynecol.+Scand%22%5BJournal%5D)">Histologic confirmation of endometriosis in different peritoneal lesions.</a> Acta. Obstet. Gynecol. Scand. 71, 337-342 (1992).</li>
<li>Cummings, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Methoxychlor+as+a+model+for+environmental+estrogens%5BTitle%5D)+AND+%22Crit.+Rev.+Toxicol%22%5BJournal%5D)">Methoxychlor as a model for environmental estrogens.</a> Crit. Rev. Toxicol. 27, 367-379 (1997).</li>
<li>Fowler, R. E., Edwards, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Induction+of+superovulation+and+pregnancy+in+mature+mice+by+gonadotrophins%5BTitle%5D)+AND+%22J.+Endocrinol%22%5BJournal%5D)">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/pubmed?term=((Comparison+of+superovulation+in+the+immature+mouse+and+rat%5BTitle%5D)+AND+%22J.+Reprod.+Fertil%22%5BJournal%5D)">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/pubmed?term=((Experimental+murine+endometriosis+induces+DNA+methylation+and+altered+gene+expression+in+eutopic+endometrium%5BTitle%5D)+AND+%22Biol.+Reprod%22%5BJournal%5D)">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/pubmed?term=((Endometrial+ability+to+implant+in+ectopic+sites+can+be+prevented+by+interleukin-12+in+a+murine+model+of+endometriosis%5BTitle%5D)+AND+%22Hum.+Reprod%22%5BJournal%5D)">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/pubmed?term=((Development+of+an+experimental+model+of+endometriosis+using+mice+that+ubiquitously+express+green+fluorescent+protein%5BTitle%5D)+AND+%22Hum.+Reprod%22%5BJournal%5D)">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/pubmed?term=((Animal+studies+in+endometriosis%3A+a+review%5BTitle%5D)+AND+%22Ilar.+J%22%5BJournal%5D)">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/pubmed?term=((Effect+of+prenatal+exposure+to+TCDD+on+the+promotion+of+endometriotic+lesion+growth+by+TCDD+in+adult+female+rats+and+mice%5BTitle%5D)+AND+%22Toxicol.+Sci%22%5BJournal%5D)">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/pubmed?term=((Effects+of+estrogen%2C+progesterone%2C+and+methoxychlor+on+surgically+induced+endometriosis+in+rats%5BTitle%5D)+AND+%22Fundam.+Appl.+Toxicol%22%5BJournal%5D)">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/pubmed?term=((Endometriotic+lesions+synthesize+and+secrete+a+haptoglobin-like+protein%5BTitle%5D)+AND+%22Biol.+Reprod%22%5BJournal%5D)">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/pubmed?term=((Genistein+causes+regression+of+endometriotic+implants+in+the+rat+model%5BTitle%5D)+AND+%22Fertil.+Steril%22%5BJournal%5D)">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/pubmed?term=((Endocannabinoid+involvement+in+endometriosis%5BTitle%5D)+AND+%22Pain%22%5BJournal%5D)">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/pubmed?term=((Nonsteroidal+antiinflammatory+drugs+differentially+suppress+endometriosis+in+a+murine+model%5BTitle%5D)+AND+%22Fertil.+Steril%22%5BJournal%5D)">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/pubmed?term=((Endostatin+inhibits+the+growth+of+endometriotic+lesions+but+does+not+affect+fertility%5BTitle%5D)+AND+%22Fertil.+Steril%22%5BJournal%5D)">Endostatin inhibits the growth of endometriotic lesions but does not affect fertility.</a> Fertil. Steril. 84, Suppl 2. 1144-1155 (2005).</li>
<li>Becker, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Short+synthetic+endostatin+peptides+inhibit+endothelial+migration+in+vitro+and+endometriosis+in+a+mouse+model%5BTitle%5D)+AND+%22Fertil.+Steril%22%5BJournal%5D)">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/pubmed?term=((Using+rats+as+a+research+model+for+the+study+of+endometriosis%5BTitle%5D)+AND+%22Ann.+N.Y.+Acad.+Sci%22%5BJournal%5D)">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/pubmed?term=((Reduced+Fecundity+in+Female+Rats+with+Surgically+Induced+Endometriosis+and+in+Their+Daughters%3A+A+Potential+Role+for+Tissue+Inhibitors+of+Metalloproteinase+1%5BTitle%5D)+AND+%22Biol.+Reprod%22%5BJournal%5D)">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>

No conflicts of interest declared.

There are several critical parameters that should be noted while performing the surgical induction of endometriosis in mice. First, endometriosis is an estrogen dependent disease and as such this surgery should be performed in intact animals or alternatively in ovariectomized animals supplemented with estrogens20. Second, suturing the endometrial biopsies to the arterial cascade must be performed with extreme care. We have found that using only two relatively loose knots with one throw each keeps the biopsy in...

<table><tbody><tr><td>Wax pencil</td><td>Fisher Scientific</td><td>NC9954135</td><td></td></tr><tr><td>Glass slide</td><td>Fisher Scientific</td><td>12-550-433</td><td></td></tr><tr><td>Eyedropper</td><td>Fisher Scientific</td><td>S79383</td><td></td></tr><tr><td>Standard light microscope for evaluating vaginal cytology smears</td><td></td><td></td><td></td></tr><tr><td>Buprenorphine HCL c3 (CARJET) 10X1ml</td><td>Butler Animal Health Supply</td><td>022891</td><td></td></tr><tr><td>Sterile phosphate buffered saline (PBS)</td><td>GIBCO, by Life Technologies</td><td>14040-117</td><td></td></tr><tr><td>10,000U/ml Penicillin, 10,000&amp;mu;g/ml Streptomycin in 0.85% NaCl</td><td>Hyclone</td><td>SV30010</td><td></td></tr><tr><td>Isoflurane</td><td>Abbott Laboratories</td><td>05260-05</td><td></td></tr><tr><td>Isoflurane non-rebreathing anesthetic system</td><td></td><td></td><td></td></tr><tr><td>Recirculating hot water heating pad</td><td></td><td></td><td></td></tr><tr><td>30 ml syringe sheath</td><td>Fisher Scientific</td><td>14-823-16G</td><td></td></tr><tr><td>Powder free sterile gloves</td><td>Fisher Scientific</td><td>19020558</td><td></td></tr><tr><td>Ophthalmic ointment</td><td>Major Pharmaceuticals</td><td>10033691</td><td></td></tr><tr><td>Small electrical clippers</td><td>Wahl Clipper Corp.</td><td>9861-600</td><td></td></tr><tr><td>Chlorhexidine scrub</td><td>Fisher Scientific</td><td>NC9863042</td><td></td></tr><tr><td>70% Ethanol</td><td></td><td></td><td></td></tr><tr><td>Polylined sterile field</td><td>Busse Hospital Disposables</td><td>696</td><td></td></tr><tr><td>Size 3 scalpel</td><td>Fisher Scientific</td><td>22-079-657</td><td></td></tr><tr><td>Number 10 scalpel blades</td><td>Fisher Scientific</td><td>22-079-681</td><td></td></tr><tr><td>Small surgical scissors</td><td>Roboz Surgical Instruments Co.</td><td>RS-5850</td><td></td></tr><tr><td>Small serrated semi-curved forceps</td><td>Roboz Surgical Instruments Co.</td><td>RS-5135</td><td></td></tr><tr><td>5-0 black braided silk suture</td><td>Ethicon Inc.</td><td>K870H</td><td></td></tr><tr><td>Sterilized pyrex glass Petri dishes</td><td>Corning</td><td>70160-101</td><td></td></tr><tr><td>2 mm biopsy punch</td><td>Miltex Inc.</td><td>33-31</td><td></td></tr><tr><td>Sterile gauze</td><td>Kendall</td><td>1806</td><td></td></tr><tr><td>6-0 black monofilament ethilon nylon suture</td><td>Ethicon Inc.</td><td>697G</td><td></td></tr><tr><td>Needle drivers (optional)</td><td>World Precision Instruments, Inc.</td><td>500023</td><td></td></tr><tr><td>5-0 undyed braided coated vicryl suture</td><td>Ethicon Inc.</td><td>J490G</td><td></td></tr><tr><td>9mm Autoclip wound clips</td><td>BD Biosciences</td><td>427631</td><td></td></tr><tr><td>Autoclip applier &amp;amp; remover</td><td>BD Biosciences</td><td>427630</td><td></td></tr><tr><td>23G needle</td><td>BD Biosciences</td><td>305193</td><td></td></tr><tr><td>1cc syringe</td><td>BD Biosciences</td><td>301025</td><td></td></tr><tr><td>5X magnifying glass stand (optional)</td><td>Fisher Scientific</td><td>14-648-23</td><td></td></tr><tr><td>10% Buffered formalin</td><td>Fisher Scientific</td><td>SF100-4</td><td></td></tr><tr><td>Calipers</td><td>Roboz Surgical Instruments Co.</td><td>RS-6466</td><td></td></tr><tr><td>Processing/embedding cassettes</td><td>Fisher Scientific</td><td>15-197-700A</td><td></td></tr><tr><td>Biopsy foam pads</td><td>Fisher Scientific</td><td>22-038-222</td><td></td></tr><tr><td>RNAqueous RNA isolation kit</td><td>Ambion</td><td>AM1912</td><td></td></tr><tr><td>Liquid nitrogen</td><td></td><td></td><td></td></tr><tr><td>Snap cap microcentrifuge flat top tube</td><td>Fisher Scientific</td><td>02-681-240</td><td></td></tr><tr><td>Ketamine (optional)</td><td>Sigma-Aldrich</td><td>K4138</td><td></td></tr><tr><td>Domitor (medetomidine hydrochloride) (optional)</td><td>Tocris Bioscience</td><td>2023</td><td></td></tr><tr><td>Antisedan (atipamezole) (optional)</td><td>Sigma-Aldrich</td><td>A9611</td><td></td></tr></tbody></table>

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

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

Endometriosis is a chronic, painful disease whose etiology remains unknown. Furthermore, treatment of endometriosis can require laparoscopic removal of lesions, and/or chronic pharmaceutical management of pain and infertility symptoms. The cost associated with endometriosis has been estimated at 22 billion dollars per year in the United States1. To further our understanding of mechanisms underlying this enigmatic disease, animal models have been employed. Primates spontaneously develop endometriosis and therefore primate models most closely resemble the disease in women. Rodent models, however, are more cost effective and readily available2. The model that we describe here involves an autologous transfer of uterine tissue to the intestinal mesentery (Figure 1) and was first developed in the rat3 and later transferred to the mouse4. The goal of the autologous rodent model of surgically-induced endometriosis is to mimic the disease in women. We and others have previously shown that the altered gene expression pattern observed in endometriotic lesions from mice or rats mirrors that observed in women with the disease5,6. One advantage of performing the surgery in the mouse is that the abundance of transgenic mouse strains available can aid researchers in determining the role of specific components important in the establishment and growth of endometriosis. An alternative model in which excised human endometrial fragments are introduced to the peritoneum of immunocompromised mice is also widely used but is limited by the lack of a normal immune system which is thought to be important in endometriosis2,7. Importantly, the mouse model of surgically induced endometriosis is a versatile model that has been used to study how the immune system8, hormones9,10 and environmental factors11,12 affect endometriosis as well as the effects of endometriosis on fertility13 and pain14.

A description of the surgical induction of endometriosis in mice and rats by auto-transplantation of uterine tissue to the arterial cascade of the intestinal mesentery.

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

The overall goal of this procedure is to surgically induce endometriosis in mice by autologous transfer of uterine tissue to the arterial cascade of the intestinal mesentary. This is accomplished by first ligating, removing and exposing the endometrium of the uterus. The second step of the procedure is to create three endometrioid implants from the exposed endometrium using a two millimeter biopsy punch.The third step of the procedure is to suture the three endometrioid implants to arteries in the arterial cascade of the intestinal mesentery. The final step of the procedure is to close the abdominal wall and skin and allow the animal to recover the endometrioid lesions. Excised from mice resemble those from women with endometriosis as determined by histological analysis and gene and protein expression patterns.Generally, individuals new to this method will struggle for three main reasons. First, the mouse tissues are just very small. Second, the intestinal mesentery is easily damaged, and third, sutures if placed too tightly, will result in bowel necrosis.Begin by gathering all the necessary surgical equipment for a successful aseptic surgery, including solutions for both analgesia and anesthesia. After anesthetizing the animal using inhaled isoflurane, confirm full anesthesia by negative response to a toe pinch stimulus to prevent dryness during surgery, apply ophthalmic ointment to the eyes and then shave the surgery site using small clippers. Disinfect the surgery site with three alternating swipes of chlorhexidine scrub and 70%ethanol, and then drape the animal with a sterile field.Make a small midline incision using either small scissors or a scalpel blade ending 0.5 to 1.0 centimeters rostral to the vaginal opening. Gently blunt, dissect the body wall from the abdominal wall in order to facilitate closure of the surgery site. After blunt dissecting, make a small midline incision in the abdominal wall.Next, using small forceps, gently locate the left uterine horn. The uterus is dorsal to the intestine, which is what is typically visible upon first entering the incision site, gently pull up on the uterine horn and slide an open forceps underneath it to serve as a retractor. It is also helpful to note the appearance of the ovaries and uterus at this time.To assist with estrus cycle dating. Slide two, six to eight centimeter pieces of five aught black braided silk suture underneath the stretched uterine horn. Securely ligate the horn at the utero tubule junction.Just coddle to the fallopian tube and at the utero cervical junction. Using a square knot at each location, leave the ends of the suture uncut. Cut out the section of uterine horn between the two ligations and place the tissue in a glass Petri dish containing approximately 100 microliters of PBS containing penicillin and streptomycin.Cut the ends of the silk suture Last. If suture comes loose or there is bleeding, find the stump and tie another knot or not using small scissors. Strip the fat off the uterine horn.Open the uterine horn by inserting one blade of a small pair of scissors into the lumen. Gently slide the scissors down the uterine horn while holding the horn with forceps In the glass Petri dish, use a two millimeter biopsy punch to cut out three equal sized implants. Being sure to keep them from drying out.Place sterile gauze immediately above the incision site and thoroughly wet with sterile PBS containing penicillin and streptomycin. It is important to maintain hydration throughout the following steps with small, smooth forceps. Gently find the cecum and move roly along the small intestine.Pull out a small four to five centimeter section of intestine and arrange it like a fan on the pre wetted gauze so that the arterial cascade of the intestinal mesentery is clearly visible. Be sure to keep the bowel moist at all times with sterile saline. Place one implant on a needle, then use six aut black alon suture to gently secure the implant to an artery approximately 0.5 centimeters from the bowel.Use only one. Throw over the forceps and do not tighten the suture very hard. As doing so may result in loss of blood flow and subsequent necrosis of the intestine.Complete two knots with one, throw each and trim the suture within two millimeters of the implant, hydrate the intestine. Then moving in a rostral direction. Pull out the next three to four centimeters of intestine and gently replace the section that already contains an implant.Skip one or two arteries from the previous implant site and suture the next implant repeat for the third implant. Finally, suture the abdominal wall and close the skin with wound clips. After closing the surgery site and administering analgesics, place the animal ventral side down in cage, partially atop a recirculating heated water pad until the animal is recovered.To recover the endometrioid lesions, locate the black sutures in the intestinal mesentery. Measure the length and width of each endometrioid lesion using calipers. Dissect out the endometrioid lesions, being careful not to lance them.Remove any non endometrioid tissue from the lesion. Wave the three fluid-filled endometrioid lesions prior to removing the suture. Gently remove the suture and lance two of the endometrioid lesions.Weigh these again before proceeding onto tissue processing. Next excise and weigh the remaining uterine horn histological analysis of endometriosis in both women and in the mouse model shown here indicates that the endometrioid lesion contains endometrial glands and stroma endometriotic lesions in mice also contain hemosiderin laden macrophages, which are a common hallmark of endometriosis in women. Endometrioid lesions removed from mice three days post induction appear inflamed and hemorrhagic as shown on the left after two to four weeks of growth.Endometriotic lesions in the mouse model are cyst like fluid-filled and surrounded by peritoneal adhesions compared to lesion weight at induction. Fluid-filled lesions were 306%and 862%larger and lance lesions were 51%and 172%larger at one and two months post induction respectively. Fluid-filled and lad endometrioid lesion.Weights remain consistent at one month post induction over five different experiments Following this procedure. This model can be used to answer questions about the pathophysiology of endometriosis and to explore novel therapeutics for the treatment of pain and infertility associated with this disease.

mouse-model-surgically-induced-endometriosis-auto-transplantation

Research

JoVE Journal

Medicine

This video demonstrates the generation of a rabbit model of endometrial cancer by injecting a highly metastatic VX2 cell suspension into the sutured uterine horns of the rabbit. This leads to generation of primary endometrial cancer with retroperitoneal lymph node metastases.

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

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...

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

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.

A Syngeneic Murine Model of Endometriosis using Naturally Cycling Mice

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.

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

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.

Establishing a Mouse Model of Thin Endometrium

This protocol describes a procedure for inducing uterine injury in a mouse model, followed by downstream analyses during a subsequent pregnancy.

Mouse Model of Surgical Uterine Injury and Subsequent Pregnancy Outcomes

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.

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

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.

Implantation of Inferior Vena Cava Interposition Graft in Mouse Model

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.

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

Here, we describe a semi-invasive optical microscopy approach for the induction of a Rose Bengal photothrombotic clot in the somatosensory cortex of a mouse in vivo.  The technical aspects of the imaging procedure are described from induction of a photothrombotic event to application and data collection.

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

Learn to surgically induce endometriosis in mice by auto-transplanting uterine tissue to the intestinal mesentery. This model creates endometrioid lesions resembling human disease for studying pathophysiology and therapeutics.

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

Explore More Videos