Name: surgical techniques to optimize ovarian reserve during laparoscopic cystectomy for ovarian endometrioma
Uploaded: 2022-01-22T13:00:00
Description: Watch this Scientific Journal Video about Surgical Techniques to Optimize Ovarian Reserve during Laparoscopic Cystectomy for Ovarian Endometrioma at JoVE.com

The patient described provided written informed consent for the use and publication of medical data, operative video, and related images for educational and scientific purposes. The following protocol adheres to the human research ethics committee guidelines of Montefiore Medical Center and New York City Health and Hospitals Corporation.
Due to limitations of the available raw recording, not every step in the protocol could be fully documented in the video on this patient.
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<ol>
	<li>Dunselman, G. A. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ESHRE+guideline:+Management+of+women+with+endometriosis.">ESHRE guideline: Management of women with endometriosis.</a> Human Reproduction. 29 (3), 400-412 (2014).</li><li>Chapron, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+management+of+deeply+infiltrating+endometriosis:+an+update.">Surgical management of deeply infiltrating endometriosis: an update.</a> Annals of the New York Academy of Sciences. 1034, 326-337 (2004).</li><li>Macer, M. L., 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=Endometriosis+and+infertility.+A+review+of+the+pathogenesis+and+treatment+of+endometriosis-associated+infertility.">Endometriosis and infertility. A review of the pathogenesis and treatment of endometriosis-associated infertility.</a> Obstetrics and Gynecology Clinics of North America. 39 (4), 535-549 (2012).</li><li>Schenken, R. S., Asch, R. H., Williams, R. F., Hodgen, G. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Etiology+of+infertility+in+monkeys+with+endometriosis:+Measurement+of+peritoneal+fluid+prostaglandins.">Etiology of infertility in monkeys with endometriosis: Measurement of peritoneal fluid prostaglandins.</a> American Journal of Obstetrics and Gynecology. 150 (4), (1984).</li><li>Raffi, F., Metwally, M., Amer, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+excision+of+ovarian+endometrioma+on+ovarian+reserve:+A+systematic+review+and+meta-analysis.">The impact of excision of ovarian endometrioma on ovarian reserve: A systematic review and meta-analysis.</a> Journal of Clinical Endocrinology and Metabolism. 97 (9), 3146-3154 (2012).</li><li>Kasapoglu, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endometrioma-related+reduction+in+ovarian+reserve+(ERROR):+a+prospective+longitudinal+study.">Endometrioma-related reduction in ovarian reserve (ERROR): a prospective longitudinal study.</a> Fertility and Sterility. 110 (1), 122-127 (2018).</li><li>Toya, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Moderate+and+severe+endometriosis+is+associated+with+alterations+in+the+cell+cycle+of+granulosa+cells+in+patients+undergoing+in+vitro+fertilization+and+embryo+transfer.">Moderate and severe endometriosis is associated with alterations in the cell cycle of granulosa cells in patients undergoing in vitro fertilization and embryo transfer.</a> Fertility and Sterility. 73 (2), (2000).</li><li>Pal, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+varying+stages+of+endometriosis+on+the+outcome+of+in+vitro+fertilization-embryo+transfer.">Impact of varying stages of endometriosis on the outcome of in vitro fertilization-embryo transfer.</a> Journal of Assisted Reproduction and Genetics. 15 (1), (1998).</li><li>Hornstein, M. D., Barbieri, R. L., McShane, P. M. <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+previous+ovarian+surgery+on+the+follicular+response+to+ovulation+induction+in+an+in+vitro+fertilization+program.">Effects of previous ovarian surgery on the follicular response to ovulation induction in an in vitro fertilization program.</a> Journal of Reproductive Medicine for the Obstetrician and Gynecologist. 34 (4), (1989).</li><li>Morcos, R. N., Gibbons, W. E., Findley, W. E. <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+peritoneal+fluid+on+in+vitro+cleavage+of+2-cell+mouse+embryos:+Possible+role+in+infertility+associated+with+endometriosis.">Effect of peritoneal fluid on in vitro cleavage of 2-cell mouse embryos: Possible role in infertility associated with endometriosis.</a> Fertility and Sterility. 44 (5), (1985).</li><li>Holoch, K. J., Lessey, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endometriosis+and+infertility.">Endometriosis and infertility.</a> Clinical Obstetrics and Gynecology. 53 (2), 429-438 (2010).</li><li>Younis, J. S., Shapso, N., Fleming, R., Ben-Shlomo, I., Izhaki, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+unilateral+versus+bilateral+ovarian+endometriotic+cystectomy+on+ovarian+reserve:+A+systematic+review+and+meta-analysis.">Impact of unilateral versus bilateral ovarian endometriotic cystectomy on ovarian reserve: A systematic review and meta-analysis.</a> Human Reproduction Update. 25 (3), 375-391 (2019).</li><li>Busacca, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Postsurgical+ovarian+failure+after+laparoscopic+excision+of+bilateral+endometriomas.">Postsurgical ovarian failure after laparoscopic excision of bilateral endometriomas.</a> American Journal of Obstetrics and Gynecology. 195 (2), 421-425 (2006).</li><li>Tsolakidis, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+on+ovarian+reserve+after+laparoscopic+ovarian+cystectomy+versus+three-stage+management+in+patients+with+endometriomas:+a+prospective+randomized+study.">The impact on ovarian reserve after laparoscopic ovarian cystectomy versus three-stage management in patients with endometriomas: a prospective randomized study.</a> Fertility and Sterility. 94 (1), 71-77 (2010).</li><li>Tsoumpou, I., Kyrgiou, M., Gelbaya, T. A., Nardo, L. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+surgical+treatment+for+endometrioma+on+in+vitro+fertilization+outcomes:+a+systematic+review+and+meta-analysis.">The effect of surgical treatment for endometrioma on in vitro fertilization outcomes: a systematic review and meta-analysis.</a> Fertility and Sterility. 92 (1), 75-87 (2009).</li><li>Medeiros, L. R. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoscopy+versus+laparotomy+for+benign+ovarian+tumour.">Laparoscopy versus laparotomy for benign ovarian tumour.</a> Cochrane Database of Systematic Reviews. (2), (2009).</li><li>Jacobson, T. Z., Duffy, J. M. N., Barlow, D., Koninckx, P. R., Garry, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoscopic+surgery+for+pelvic+pain+associated+with+endometriosis.+[Review]+[36+refs][Update+in+Cochrane+Database+Syst+Rev.+2014;8:CD001300;+PMID:+25130257],+[Update+of+Cochrane+Database+Syst+Rev.+2001;(4):CD001300;+PMID:+11687104].">Laparoscopic surgery for pelvic pain associated with endometriosis. [Review] [36 refs][Update in Cochrane Database Syst Rev. 2014;8:CD001300; PMID: 25130257], [Update of Cochrane Database Syst Rev. 2001;(4):CD001300; PMID: 11687104].</a> The Cochrane Database of Systematic Reviews. , (2009).</li><li>Carmona, F., Mart&#237;nez-Zamora, M. A., Rabanal, A., Mart&#237;nez-Rom&#225;n, S., Balasch, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ovarian+cystectomy+versus+laser+vaporization+in+the+treatment+of+ovarian+endometriomas:+A+randomized+clinical+trial+with+a+five-year+follow-up.">Ovarian cystectomy versus laser vaporization in the treatment of ovarian endometriomas: A randomized clinical trial with a five-year follow-up.</a> Fertility and Sterility. 96 (1), (2011).</li><li>Hart, R. J., Hickey, M., Maouris, P., Buckett, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Excisional+surgery+versus+ablative+surgery+for+ovarian+endometriomata.">Excisional surgery versus ablative surgery for ovarian endometriomata.</a> Cochrane Database of Systematic Reviews. (2), (2008).</li><li>Vignali, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+excision+of+ovarian+endometriomas:+Does+it+truly+impair+ovarian+reserve?+Long+term+anti-M&#252;llerian+hormone+(AMH)+changes+after+surgery.">Surgical excision of ovarian endometriomas: Does it truly impair ovarian reserve? Long term anti-M&#252;llerian hormone (AMH) changes after surgery.</a> Journal of Obstetrics and Gynaecology Research. 41 (11), 1773-1778 (2015).</li><li>Revised American Society for Reproductive Medicine. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Revised+American+Society+for+Reproductive+Medicine+classification+of+endometriosis.">Revised American Society for Reproductive Medicine classification of endometriosis.</a> Fertility and Sterility. 67 (5), 817-821 (1997).</li><li>Adamson, G. D., Pasta, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endometriosis+fertility+index:+the+new,+validated+endometriosis+staging+system.">Endometriosis fertility index: the new, validated endometriosis staging system.</a> Fertility and Sterility. 94 (5), 1609-1615 (2010).</li><li>Endometriosis and infertility Committee. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endometriosis+and+infertility:+A+committee+opinion.">Endometriosis and infertility: A committee opinion.</a> Fertility and Sterility. 98 (3), (2012).</li><li>Miller, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+endometrioma+treatment+paradigm+when+fertility+is+desired:+A+systematic+review.">The endometrioma treatment paradigm when fertility is desired: A systematic review.</a> Journal of Minimally Invasive Gynecology. 28 (3), 575-586 (2021).</li><li>Muzii, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Histologic+analysis+of+specimens+from+laparoscopic+endometrioma+excision+performed+by+different+surgeons:+Does+the+surgeon+matter.">Histologic analysis of specimens from laparoscopic endometrioma excision performed by different surgeons: Does the surgeon matter.</a> Fertility and Sterility. 95 (6), (2011).</li><li>Kotlyar, A., Gingold, J., Shue, S., Falcone, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+salpingectomy+on+ovarian+function.">The effect of salpingectomy on ovarian function.</a> Journal of Minimally Invasive Gynecology. 24 (4), 563-578 (2017).</li></ol>

Patients with diagnosed endometriosis commonly report pain or implant-related organ dysfunction, including infertility. Up to 50% of patients with endometriosis meet criteria for infertility23. Ovarian reserve, measured via AMH levels, FSH levels around menses or an antral follicle count, is used to predict patient response to gonadotropin stimulation. Surgical management of endometrioma is known to decrease ovarian reserve23. However, ovarian reserve continues to decline i...

Endometriosis is a chronic inflammatory condition defined by ectopic endometrial tissue1. Patients with endometriosis commonly present with pain-related complaints and organ dysfunction related to the site of ectopic implantation, which may be anywhere in the body though is typically within the pelvis2,3. Ovarian endometrioma, in which a cyst of endometriosis forms within the ovary, contributes to subfertility by multiple means, including depletion of the ovarian follicle pool, promoting an inflammatory environment, progesterone resistance, and impaired ovum release and capture4. Affected patients frequently have diminished ovarian reserve upon presentation, and the follicle pool is further depleted following surgery5,6. Cyclic bleeding in sites with ectopic endometrial tissue leads to inflammation and significant adhesions, which may impair ovum pick-up and transport, fertilization, and embryo passage7,8,9. The abnormal inflammatory environment may also impair folliculogenesis and early embryonic development10,11.
At a practical surgical level, the resultant adhesions frequently obliterate normally avascular surgical planes, placing patients at elevated risk of prolonged operative times, blood loss, and surgical injury. Surgical management of patients desiring future fertility presents additional challenges, most notably risk of iatrogenic damage to the tubes and ovaries, compromising tubal patency or ovarian reserve12,13,14. However, surgical resection of endometriosis also presents a unique opportunity to potentially diagnose, evaluate, and treat identified lesions, including optimizing fertility15.
We detail our approach to the treatment of ovarian endometriomas in patients desiring future fertility. We strongly favor a laparoscopic approach over laparotomy for enhanced post-operative results, including less pain, shorter hospital stay, and quicker recovery16. This protocol prioritizes excision of ovarian endometrioma over drainage and ablation as it provides a more favorable outcome with respect to cyst recurrence, pain recurrence, and spontaneous pregnancies17,18,19,20. Additionally, it allows for the specimen retrieval for histology. We excise ovarian endometrioma via a stripping technique with limited electrosurgical energy application. Nonetheless, many specialized tools have been described for the management of ovarian endometrioma and multiple acceptable surgical approaches exist for patients desiring future fertility. Regardless of any specific technique employed, the considerations and surgical principles described in this protocol are applicable to all patients with endometriosis desiring fertility-optimizing surgery.
The protocol described below was employed for the care of a 32-year-old woman with chronic pelvic pain and histologically proven endometriosis identified during a prior laparoscopy, during which endometriosis was incompletely resected. She endorsed pain refractory to the first-line medical management and was interested in future fertility, although had not actively attempted spontaneous conception. She was deemed a candidate for minimally invasive surgical resection and underwent an exam under anesthesia, diagnostic laparoscopy, lysis of adhesions, ovarian cystectomy, chromopertubation, and salpingectomy. Her surgery and recovery were uncomplicated.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Basic laparoscopy</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1L bag 0.9% NaCl solution</td>
			<td></td>
			<td></td>
			<td>Suction irrigation, hemostasis, lysis of adhesions</td>
		</tr>
		<tr>
			<td>1-10% povidone-iodine</td>
			<td></td>
			<td></td>
			<td>Sterile prep</td>
		</tr>
		<tr>
			<td>2% chlorhexidine gluconate in 70% isopropyl alcohol for skin</td>
			<td></td>
			<td></td>
			<td>Sterile prep</td>
		</tr>
		<tr>
			<td>4% chlorhexidine gluconate</td>
			<td></td>
			<td></td>
			<td>Sterile prep</td>
		</tr>
		<tr>
			<td>5mm laparoscopic trocar and sleeve x 3</td>
			<td>Covidien</td>
			<td>ONB5STF</td>
			<td>Diagnostic laparoscopy</td>
		</tr>
		<tr>
			<td>12mm laparoscopic trocar and sleeve</td>
			<td>Covidien</td>
			<td>ONB12STF</td>
			<td>Diagnostic laparoscopy and larger size facilitates specimen retrieval bag</td>
		</tr>
		<tr>
			<td>CO2 insufflator</td>
			<td>Stryker</td>
			<td>620-040-504</td>
			<td>Diagnostic laparoscopy</td>
		</tr>
		<tr>
			<td>CO2 insufflator tubing</td>
			<td>Stryker</td>
			<td>620-030-201</td>
			<td>Diagnostic laparoscopy</td>
		</tr>
		<tr>
			<td>Electrosurgical generator</td>
			<td>Covidien</td>
			<td>VLFT10GEN</td>
			<td>Diagnostic laparoscopy</td>
		</tr>
		<tr>
			<td>Foley kit and urometer bag</td>
			<td>Bard</td>
			<td>153214</td>
			<td>Diagnostic laparoscopy</td>
		</tr>
		<tr>
			<td>Laparoscopic scope, 5mm 0 degree</td>
			<td>Olympus</td>
			<td>WA4KL500</td>
			<td>Diagnostic laparoscopy</td>
		</tr>
		<tr>
			<td>Laparoscopic scope, 5mm 30 degree</td>
			<td>Olympus</td>
			<td>WA4KL530</td>
			<td>Diagnostic laparoscopy</td>
		</tr>
		<tr>
			<td>Laparoscopic suction irrigation pool tip</td>
			<td>Stryker</td>
			<td>250-070-406</td>
			<td>Suction irrigation, hemostasis, lysis of adhesions</td>
		</tr>
		<tr>
			<td>StrykeFlow II suction irrigator</td>
			<td>Stryker</td>
			<td>250-070-500</td>
			<td>Suction irrigation, hemostasis, lysis of adhesions</td>
		</tr>
		<tr>
			<td>Suction tubing</td>
			<td>Stryker</td>
			<td>250-070-403</td>
			<td>Suction irrigation, hemostasis, lysis of adhesions</td>
		</tr>
		<tr>
			<td>Suction/vacuum source</td>
			<td></td>
			<td></td>
			<td>Suction irrigation, hemostasis, lysis of adhesions</td>
		</tr>
		<tr>
			<td>Cystectomy and Salpingectomy</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>10mm tissue retrieval bag</td>
			<td>Covidien</td>
			<td>173050G</td>
			<td>Specimen retrieval bag to be used in 12mm laparoscopic trocar site</td>
		</tr>
		<tr>
			<td>Atraumatic laparoscopic bowel graspers</td>
			<td>Stryker</td>
			<td>250-080-319</td>
			<td>Lysis of adhesions, cystectomy, salpingectomy</td>
		</tr>
		<tr>
			<td>Bipolar Cord</td>
			<td>Stryker</td>
			<td>250-040-016</td>
			<td>Lysis of adhesions, cystectomy, salpingectomy</td>
		</tr>
		<tr>
			<td>Endopeanut laparoscopic retractor</td>
			<td>Covidien</td>
			<td>173019</td>
			<td>Lysis of adhesions, cystectomy, salpingectomy</td>
		</tr>
		<tr>
			<td>Harmonic ACE +7 ultrasonic shears</td>
			<td>Ethicon</td>
			<td>HARH23</td>
			<td>Lysis of adhesions, cystectomy, salpingectomy</td>
		</tr>
		<tr>
			<td>Laparoscopic bipolar grasping forceps</td>
			<td>Karl Storz</td>
			<td>38951 MD</td>
			<td>Lysis of adhesions, cystectomy, salpingectomy</td>
		</tr>
		<tr>
			<td>Laparoscopic curved Metzenbaum scissors</td>
			<td>Stryker</td>
			<td>250-080-267</td>
			<td>Lysis of adhesions, cystectomy, salpingectomy</td>
		</tr>
		<tr>
			<td>Ultrasonic generator unit</td>
			<td>Ethicon</td>
			<td>GEN11</td>
			<td>Lysis of adhesions, cystectomy, salpingectomy</td>
		</tr>
		<tr>
			<td>Vasopressin solution (20 units in 50 to 100cc of injectable saline)</td>
			<td></td>
			<td></td>
			<td>Cystectomy</td>
		</tr>
		<tr>
			<td>Chromopertubation</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>10mg methylene blue in 150mL 0.9% NaCl solution</td>
			<td></td>
			<td></td>
			<td>Pigmented solution for chromopertubation</td>
		</tr>
		<tr>
			<td>ZUMI uterine manipulator/injector</td>
			<td>Cooper Surgical</td>
			<td>ZSI1151</td>
			<td>Chromopertubation, diagnostic laparoscopy, lysis of adhesions</td>
		</tr>
		<tr>
			<td>Hemostatic agents</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Arista absorbable hemostatic particles 3g</td>
			<td>Bard Davol</td>
			<td>SM0002-USA</td>
			<td>Hemostatic agent</td>
		</tr>
		<tr>
			<td>Floseal gelatin thrombin matrix 5ml</td>
			<td>Baxter</td>
			<td>ADS201844</td>
			<td>Hemostatic agent</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>10mm tissue retrieval bag</td>
			<td>Covidien</td>
			<td>173050G</td>
			<td></td>
		</tr>
		<tr>
			<td>Arista absorbable hemostatic particles 3g</td>
			<td>Bard Davol</td>
			<td>SM0002-USA</td>
			<td>Hemostatic agent</td>
		</tr>
		<tr>
			<td>Atraumatic laparoscopic bowel graspers</td>
			<td>Stryker</td>
			<td>250-080-319</td>
			<td></td>
		</tr>
		<tr>
			<td>Endopeanut laparoscopic retractor</td>
			<td>Covidien</td>
			<td>173019</td>
			<td></td>
		</tr>
		<tr>
			<td>Floseal gelatin thrombin matrix 5ml</td>
			<td>Baxter</td>
			<td>ADS201844</td>
			<td>Hemostatic agent</td>
		</tr>
		<tr>
			<td>Harmonic ACE +7 ultrasonic shears</td>
			<td>Ethicon</td>
			<td>HARH23</td>
			<td></td>
		</tr>
		<tr>
			<td>Laparoscopic bipolar grasping forceps</td>
			<td>Karl Storz</td>
			<td>38951 MD</td>
			<td></td>
		</tr>
		<tr>
			<td>Laparoscopic curved Metzenbaum scissors</td>
			<td>Stryker</td>
			<td>250-080-267</td>
			<td></td>
		</tr>
		<tr>
			<td>Methylene blue 10mg in 150mL 0.9% NaCl solution</td>
			<td></td>
			<td></td>
			<td>For chromopertubation</td>
		</tr>
		<tr>
			<td>ZUMI uterine manipulator/injector</td>
			<td>Cooper Surgical</td>
			<td>ZSI1151</td>
			<td>For uterine manipulation and chromopertubation</td>
		</tr>
	</tbody>
</table>

surgical techniques to optimize ovarian reserve during laparoscopic cystectomy for ovarian endometrioma

Surgical management of ovarian endometrioma in patients desiring fertility is complicated by the need to balance maximal resection of disease with efforts to spare normal ovarian cortex. Optimization of tubal anatomy is another frequent consideration. Fertility-sparing laparoscopic techniques at the time of cystectomy for ovarian endometrioma seek to limit iatrogenic surgical damage to the ovarian cortex and strategically assess and respond to genital tract patency. Surgical candidates frequently desire relief from endometriosis-associated pain while also seeking to optimize spontaneous or assisted conception rates. Operative benefits include potential for surgical and histopathologic diagnosis of endometriosis, evaluation of genital tract patency, and treatment of visualized lesions. Resection of ovarian endometrioma nonetheless poses significant risks, including surgical injury, blood loss, post-surgical decline in ovarian reserve and post-operative inflammation with adhesion formation, both of which may impair folliculogenesis.
We present the case of a 32-year-old woman with known endometriosis and continued pain refractory to medical management who opted for surgical management of her disease tailored toward optimizing her chances at future conception. Using this case as an example, we describe techniques and considerations for diagnostic laparoscopy, adhesiolysis, ovarian cystectomy, chromopertubation, and salpingectomy with a focus on maintaining a fertility-preserving approach.

Table 1 shows results of our patient example. Total operation time was 251 min from anesthetic induction to extubation, with an estimated blood loss of 200 mL. The recovery period was uncomplicated. As she desired future pregnancy remote from the time of surgery, she began oral contraceptives. Histopathological examination revealed a right hydrosalpinx with paratubal cyst, bilateral endometriomas (4.5 cm and 3.7 cm), and ovarian tissue.
The surgery utilized a combination of fe...

Watch this Scientific Journal Video about Surgical Techniques to Optimize Ovarian Reserve during Laparoscopic Cystectomy for Ovarian Endometrioma at JoVE.com

Surgical Techniques to Optimize Ovarian Reserve during Laparoscopic Cystectomy for Ovarian Endometrioma

This protocol presents techniques to laparoscopically excise ovarian endometrioma, to perform adhesiolysis with sparing electrosurgical application, and to employ intraoperative chromopertubation to assess for genital tract patency. This systematic approach will facilitate optimal endometriosis management, guide concomitant adnexal surgeries, and enhance post-surgical fertility outcomes.

Surgical management of ovarian endometrioma in patients desiring fertility is complicated by the need to balance maximal resection of disease with efforts ...

Surgical techniques to optimize ovarian reserve during laparoscopic cystectomy for ovarian endometrioma. This protocol describes several fertility-sparing techniques performed during cystectomy for ovarian endometrioma. These techniques demonstrate ways to limit surgical damage to the ovarian tissue, allow for a full evaluation of the pelvic anatomy, and identify and treat genital tract pathology.The following is demonstrated with a patient example obtained with written informed consent for the use and publication of medical data, video, and related images for educational and scientific purposes. This protocol follows guidelines of our institution's Human Research Ethics Committee. We will discuss the case of a 32-year-old woman desiring future fertility.She has biopsy-proven endometriosis from a prior diagnostic procedure, which left her disease incompletely resected and resulted in medically refractory pain. She was a candidate for minimally invasive approach to resect endometriosis. Step one, the preoperative evaluation.Every patient should complete an in-person consultation to obtain a thorough history and physical exam. Additionally, the patient should be screened to determine which preoperative imaging study may be helpful. Transvaginal ultrasonography is a standard imaging modality, as it usually provides adequate visualization and resolution of the uterus, posterior cul-de-sac, right adnexa, and left adnexa.Inpatients with endometriosis, consider obtaining an MRI to evaluate for the presence of adnexal pathology, adenomyosis, hydronephrosis, and evidence of deep infiltrating endometriosis, which may require further surgical planning. These findings may be reliably assessed with ultrasound alone in most patients with an experienced radiologist and aid of bowel prep. Determine whether the patient is an appropriate surgical candidate.Indications for cystectomy include size greater than four centimeters with pain or infertility or size less than four centimeters with medically refractory pain or concern for follicle count or accessibility during assisted reproduction. The patient must have no other surgical contraindications. This patient had lesions up to five centimeters plus medically refractory pain and was thus a candidate for cystectomy.The patient must be informed about the potential risks and benefits of the proposed surgery. A more complete resection risks further decline in ovarian reserve. The patient should be informed of management options regarding abnormal salpinges.In cases of deep infiltrating endometriosis, significant surgical morbidity and need for bowel resection must be addressed. Following an informed consent discussion, this patient elects for a diagnostic laparoscopy, excision of endometriosis, cystectomy, chromopertubation, and possibly salpingectomy. Now we begin our demonstration of indicated operative steps during surgeries for endometrioma resection with discussion of the exam under anesthesia and diagnostic laparoscopy.After an exam under anesthesia, create a pneumoperitoneum with standard techniques, with entry either periumbilically or at Palmer's point, as denoted by U and P on the figure. Place at least two additional lower abdominal ports, as shown here in the figure. First, perform a diagnostic laparoscopy to exclude surgical injury and to identify anatomy, adhesions, and possible endometriosis implants in the peritoneal cavity.Place a uterine manipulator and utilize steep Trendelenburg position to optimize visualization of pelvic structures. Systematically evaluate all pelvic structures and spaces. For diagnostic purposes, biopsy all possible endometriotic implants.Endometriosis can be scored to reliably describe the patient's condition using either the Revised American Society for Reproductive Medicine System or the Endometriosis Fertility Index, which is most appropriate for predicting fertility outcomes. Often, adhesiolysis is required for adequate exposure. Lysis of adhesions allow for adequate tissue handling.Once port placement is optimal, prepared to dissect adhesions to expose the adnexa. Divide the kissing ovaries and lyse tubal adhesions. Start with blunt dissection from the healthy tissue proximally, working towards dense adhesions and abnormal anatomy.Triangulate applied forces to allow for efficient dissection. Atraumatic graspers can apply traction to sensitive tissues, while the suction irrigator can push, sweep, hydrodissect, and aspirate for enhanced visualization. Continue along a dissection plane until it can no longer be easily developed bluntly.Use scissors or energy devices like ultrasonic shears to dissect dense adhesions. Ensure full tubal length is freed from adhesions, including fimbriae. When working in proximity to major structures, the surgeon must be prepared to perform retroperitoneal dissection and ureterolysis for safe identification of anatomy prior to dividing adherent structures.The next step is to evaluate for genital tract patency. We demonstrate this with chromopertubation. Through the uterine manipulator, inject dilute methylene blue.Evaluate laparoscopically for distension of the fallopian tube and spillage of dye from the fimbriae. When tubal patency is not demonstrated, ensure satisfactory fimbriolysis. Next, use atraumatic graspers to occlude the patent tube proximally.This will favor flow of dye through the contralateral side. Finally, the surgeon can replace the uterine manipulator for a dedicated catheter commonly used during hysterosalpingogram to reattempt distension. After tubal pathology is identified and evaluated, consider removal of diseased portions if consistent with the surgical plan.Here, we perform a partial right salpingectomy after identification of the infundibulopelvic ligament, labeled with a star. Exposing the tube with graspers, start the dissection of the tubal lumen from the mesosalpinx at its fimbriated end. Limit the amount of mesosalpinx excise to preserve anastomotic vascular connections that supply ovarian tissue.Reaching the isthmus, transect across the full tubal lumen to complete this step. The patient should be informed of her tubal status, as well as the presence of any remaining tubal tissue after surgery. The next step is cystectomy for ovarian endometrioma.Divide remaining adhesions to free the ovary. Any disruption of the endometrioma cyst wall will yield chocolate-colored fluid. Elevate the ovary with graspers to facilitate exposure of the lesion.Incise the thinnest portion of ovarian cortex to expose the cyst wall. In this example, the endometrioma is then drained to identify the cleavage plane. Using traction and counter traction, separate the cyst wall from normal ovarian parenchyma.This step is difficult, as adherent cyst wall is often removed with normal ovarian tissue. Efficient dissection requires close placement of graspers with applied force perpendicular to the dissection plane. This will avoid tearing.In this example, optimal technique would have also used graspers placed on the ovarian side, closest to the dissection plane. After complete excision, assess the operative bed for hemostasis. Use copious irrigation and aspiration to remove hemoperitoneum and evaluate for active bleeding.Start by observing the ovary for one to three minutes as the endogenous coagulation cascade is activated. If bleeding continues, try topical hemostatic agents. Consider suturing for actively bleeding vascular lesions.In our patient's example, we applied focused ultrasonic energy as we were assured of a safe margin away from genitourinary and gastrointestinal structures. Surgeons should minimize energy application to sensitive tissues, such as the ovary and its related blood supply. Consider sources with the least potential for lateral thermal spread, such as laser vaporization with carbon dioxide, ultrasonic energy, followed by bipolar and lastly monopolar electrosurgery, which has the greatest potential for collateral damage and inadvertent injury.Once hemostasis is assured, gather all specimens for retrieval into a bag to reduce the risk of port site endometriosis. Specimen removal should be under direct visualization. Address remaining sites of endometriosis.Specific approaches and techniques for this vary, depending on the site of implantation. Peritoneal implants can be biopsied and stripped, while removal of deep disease that cannot be shaved may require coordination with specialists, such as urologic or colorectal surgeons. Confirm hemostasis at the operative bed.Fascial defects larger than one centimeter are closed, followed by skin closure. The total operative time was 251 minutes. There was a total estimated blood loss of 200 cc's.The recovery period was uncomplicated and histopathological examination revealed a right hydrosalpinx with paratubal cyst, bilateral endometrioma measuring 3.7 and 4.5 centimeters, and ovarian tissue. In conclusion, this is a detailed protocol describing the steps required to perform a laparoscopic ovarian cystectomy for endometrioma using a fertility-sparing approach. Surgical management for endometriosis is favored for diagnosis, excision of disease, which reduces medically refractory pain, recurrence, and improves spontaneous pregnancy rates.Cystectomy may be helpful in patients pursuing assisted reproductive technologies. However, excision should only be performed if the patient is expected to have adequate ovarian reserve postoperatively. The surgeon must balance satisfactory removal of ovarian pathology to treat symptoms while limiting iatrogenic damage to normal ovarian parenchyma.Tubal patency is frequently assessed and management should be tailored to the patient's reproductive plans.

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Medicine

Heterotypic Three-dimensional In Vitro Modeling of Stromal-Epithelial Interactions During Ovarian Cancer Initiation and Progression

Epithelial ovarian cancers (EOCs) are the leading cause of death from gynecological malignancy in Western societies. Despite advances in surgical treatments and improved platinum-based chemotherapies, there has been little improvement in EOC survival rates for more than four decades 1,2. Whilst stage I tumors have 5-year survival rates &gt;85%, survival rates for stage III/IV disease are &lt;40%. Thus, the high rates of mortality for EOC could be significantly decreased if tumors were detected at earlier, more treatable, stages 3-5. At present, the molecular genetic and biological basis of early stage disease development is poorly understood. More specifically, little is known about the role of the microenvironment during tumor initiation; but known risk factors for EOCs (e.g. age and parity) suggest that the microenvironment plays a key role in the early genesis of EOCs. We therefore developed three-dimensional heterotypic models of both the normal ovary and of early stage ovarian cancers. For the normal ovary, we co-cultured normal ovarian surface epithelial (IOSE) and normal stromal fibroblast (INOF)&nbsp;cells, immortalized by retrovrial transduction of the catalytic subunit of human telomerase holoenzyme (hTERT) to extend the lifespan of these cells in culture. To model the earliest stages of ovarian epithelial cell transformation, overexpression of the CMYC oncogene in IOSE cells, again co-cultured with INOF cells. These heterotypic models were used to investigate the effects of aging and senescence on the transformation and invasion of epithelial cells. Here we describe the methodological steps in development of these three-dimensional model; these methodologies aren't specific to the development of normal ovary and ovarian cancer tissues, and could be used to study other tissue types where stromal and epithelial cell interactions are a fundamental aspect of the tissue maintenance and disease development.

Heterotypic Three-dimensional In Vitro Modeling of Stromal-Epithelial Interactions During Ovarian Cancer Initiation and Progression

We describe methodologies for establishing in vitro heterotypic three-dimensional models comprising ovarian fibroblasts and normal ovarian surface or ovarian cancer epithelial cells. We discuss the use of these models to study stromal-epithelial interactions that occur during ovarian cancer development.

Epithelial ovarian cancers (EOCs) are the leading cause of death from gynecological malignancy in Western societies. Despite advances in surgical ...

Method for Obtaining Primary Ovarian Cancer Cells From Solid Specimens

Reliable tools for investigating ovarian cancer initiation and progression are urgently needed. While the use of ovarian cancer cell lines remains a valuable tool for understanding ovarian cancer, their use has many limitations. These include the lack of heterogeneity and the plethora of genetic alterations associated with extended in vitro passaging.&#160;Here we describe a method that allows for rapid establishment of primary ovarian cancer cells form solid clinical specimens collected at the time of surgery. The method consists of subjecting clinical specimens to enzymatic digestion for 30 min. The isolated cell suspension is allowed to grow and can be used for downstream application including drug screening. The advantage of primary ovarian cancer cell lines over established ovarian cancer cell lines is that they are representative of the original specific clinical specimens they are derived from and can be derived from different sites whether primary or metastatic ovarian cancer.

This study describes a detailed method for isolation and characterization of primary ovarian cancer cells from solid clinical specimens. Ovarian cancer clinical specimens are subjected to enzymatic digestion to obtain viable, fibroblast-free epithelial ovarian cancer (EOC) cells highly suitable for downstream applications.

Reliable tools for investigating ovarian cancer initiation and progression are urgently needed. While the use of ovarian cancer cell lines remains a ...

Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of care consisting of surgical debulking and combination chemotherapy consisting of platinum and taxane compounds, almost 90% of patients recur within a few years. In these patients the development of chemoresistant disease limits the efficacy of currently available chemotherapy agents and therefore contributes to the high mortality. To discover novel therapy options that can target recurrent disease, appropriate animal models that closely mimic the clinical profile of patients with recurrent ovarian cancer are required. The challenge in monitoring intra-peritoneal (i.p.) disease limits the use of i.p. models and thus most xenografts are established subcutaneously. We have developed a sensitive optical imaging platform that allows the detection and anatomical location of i.p. tumor mass. The platform includes the use of optical reporters that extend from the visible light range to near infrared, which in combination with 2-dimensional X-ray co-registration can provide anatomical location of molecular signals. Detection is significantly improved by the use of a rotation system that drives the animal to multiple angular positions for 360 degree imaging, allowing the identification of tumors that are not visible in single orientation. This platform provides a unique model to non-invasively monitor tumor growth and evaluate the efficacy of new therapies for the prevention or treatment of recurrent ovarian cancer.

We describe a non-invasive animal imaging platform that allows the detection, quantification, and monitoring of ovarian cancer growth and recurrence. This intra-peritoneal xenograft model mimics the clinical profile of patients with ovarian cancer.

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of ...

Enrichment for Chemoresistant Ovarian Cancer Stem Cells from Human Cell Lines

Cancer stem cells (CSCs) are defined as a subset of slow cycling and undifferentiated cells that divide asymmetrically to generate highly proliferative, invasive, and chemoresistant tumor cells. Therefore, CSCs are an attractive population of cells to target therapeutically. CSCs are predicted to contribute to a number of types of malignancies including those in the blood, brain, lung, gastrointestinal tract, prostate, and ovary. Isolating and enriching a tumor cell population for CSCs will enable researchers to study the properties, genetics, and therapeutic response of CSCs. We generated a protocol that reproducibly enriches for ovarian cancer CSCs from ovarian cancer cell lines (SKOV3 and OVCA429). Cell lines are treated with 20 &#181;M cisplatin for 3 days. Surviving cells are isolated and cultured in a serum-free stem cell media containing cytokines and growth factors. We demonstrate an enrichment of these purified CSCs by analyzing the isolated cells for known stem cell markers Oct4, Nanog, and Prom1 (CD133) and cell surface expression of CD177 and CD133. The CSCs exhibit increased chemoresistance. This method for isolation of CSCs is a useful tool for studying the role of CSCs in chemoresistance and tumor relapse.

Cancer stem cells (CSCs) are&#160;implicated in tumor relapse due to chemoresistance. We have optimized a protocol for selection and expansion of CSCs from ovarian cancer cell lines. By treating cells with the chemotherapeutic cisplatin and culturing&#160;in a stem cell promoting media we enrich for non-adherent CSC cultures.

Cancer stem cells (CSCs) are defined as a subset of slow cycling and undifferentiated cells that divide asymmetrically to generate highly proliferative, ...

In vitro Enrichment of Ovarian Cancer Tumor-initiating Cells

Evidence suggests that small subpopulations of tumor cells maintain a unique self-renewing and differentiation capacity and may be responsible for tumor initiation and/or relapse. Clarifying the mechanisms by which these tumor-initiating cells (TICs) support tumor formation and progression could lead to the development of clinically favorable therapies. Ovarian cancer is a heterogeneous and highly recurrent disease. Recent studies suggest TICs may play an important role in disease biology. We have identified culture conditions that enrich for TICs from ovarian cancer cell lines. Growing either adherent cells or non-adherent &#8216;floater&#8217; cells in a low attachment plate with serum free media in the presence of growth factors supports the propagation of ovarian cancer TICs with stem cell markers (CD133 and ALDH activity) and increased tumorigenicity without the need to physically separate the TICs from other cell types within the culture. Although the presence of floater cells is not common for all cell lines, this population of cells with innate low adherence may have high tumorigenic potential.Compared to adherent cells grown in the presence of serum, TICs readily form spheres, are significantly more tumorigenic in mice, and express putative stem cell markers. The conditions are easy to establish in a timely manner and can be used to study signaling pathways important for maintaining stem characteristics, and to identify drugs or combinations of drugs targeting TICs. The culture conditions described herein are applicable for a variety of ovarian cancer cells of epithelial origin and will be critical in providing new information about the role of TICs in tumor initiation, progression, and relapse.

In vitro Enrichment of Ovarian Cancer Tumor-initiating Cells

Tumor-initiating cells (TICs) may represent a viable therapeutic target for the treatment of ovarian cancer, a highly recurrent and fatal disease. We present a protocol for culture conditions that enrich for this highly tumorigenic population of cells.

Evidence suggests that small subpopulations of tumor cells maintain a unique self-renewing and differentiation capacity and may be responsible for tumor ...

In Vivo and Ex Vivo Approaches to Study Ovarian Cancer Metastatic Colonization of Milky Spot Structures in Peritoneal Adipose

High-grade serous ovarian cancer (HGSC), the cause of widespread peritoneal metastases, continues to have an extremely poor prognosis; fewer than 30% of women are alive 5 years after diagnosis. The omentum is a preferred site of HGSC metastasis formation. Despite the clinical importance of this microenvironment, the contribution of omental adipose tissue to ovarian cancer progression remains understudied. Omental adipose is unusual in that it contains structures known as milky spots, which are comprised of B, T, and NK cells, macrophages, and progenitor cells surrounding dense nests of vasculature. Milky spots play a key role in the physiologic functions of the omentum, which are required for peritoneal homeostasis. We have shown that milky spots also promote ovarian cancer metastatic colonization of peritoneal adipose, a key step in the development of peritoneal metastases. Here we describe the approaches we developed to evaluate and quantify milky spots in peritoneal adipose and study their functional contribution to ovarian cancer cell metastatic colonization of omental tissues both in vivo and ex vivo. These approaches are generalizable to additional mouse models and cell lines, thus enabling the study of ovarian cancer metastasis formation from initial localization of cells to milky spot structures to the development of widespread peritoneal metastases.

In Vivo and Ex Vivo Approaches to Study Ovarian Cancer Metastatic Colonization of Milky Spot Structures in Peritoneal Adipose

We outline a protocol that implements both in vivo and ex vivo approaches to study ovarian cancer colonization of peritoneal adipose tissues, particularly the omentum. Furthermore, we present a protocol to quantitate and analyze immune cell-structures in the omentum known as milky spots, which promote metastases of peritoneal adipose.

High-grade serous ovarian cancer (HGSC), the cause of widespread peritoneal metastases, continues to have an extremely poor prognosis; fewer than 30% of ...

Quantitation of Intra-peritoneal Ovarian Cancer Metastasis

Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancy in the United States. Mortality is due to diagnosis of 75% of women with late stage disease, when metastasis is already present. EOC is characterized by diffuse and widely disseminated intra-peritoneal metastasis. Cells shed from the primary tumor anchor in the mesothelium that lines the peritoneal cavity as well as in the omentum, resulting in multi-focal metastasis, often in the presence of peritoneal ascites. Efforts in our laboratory are directed at a more detailed understanding of factors that regulate EOC metastatic success. However, quantifying metastatic tumor burden represents a significant technical challenge due to the large number, small size and broad distribution of lesions throughout the peritoneum. Herein we describe a method for analysis of EOC metastasis using cells labeled with red fluorescent protein (RFP) coupled with in vivo multispectral imaging. Following intra-peritoneal injection of RFP-labelled tumor cells, mice are imaged weekly until time of sacrifice. At this time, the peritoneal cavity is surgically exposed and organs are imaged in situ. Dissected organs are then placed on a labeled transparent template and imaged ex vivo. Removal of tissue auto-fluorescence during image processing using multispectral unmixing enables accurate quantitation of relative tumor burden. This method has utility in a variety of applications including therapeutic studies to evaluate compounds that may inhibit metastasis and thereby improve overall survival.

Ovarian cancer metastasis is characterized by numerous diffuse intra-peritoneal lesions, such that accurate visual quantitation of tumor burden is challenging. Herein we describe a method for in situ and ex vivo quantitation of metastatic tumor burden using red fluorescent protein (RFP)-labeled tumor cells and optical imaging.

Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancy in the United States.  Mortality is due to diagnosis of 75% of ...

Transplantation Into the Mouse Ovarian Fat Pad

Orthotopic transplantation assays in mice are invaluable for studies of cell regeneration and neoplastic transformation. Common approaches for orthotopic transplantation of ovarian surface and tubal epithelia include intraperitoneal and intrabursal administration of cells. The respective limitations of these methods include poorly defined location of injected cells and limited space volume. Furthermore, they are poorly suited for long-term structural preservation of transplanted organs. To address these challenges, we have developed an alternative approach, which is based on the introduction of cells and tissue fragments into the mouse fat pad. The mouse ovarian fat pad is located in the immediate vicinity of the ovary and uterine tube (aka oviduct, fallopian tube), and provides a familiar microenvironment for cells and tissues of these organs. In our approach fluorescence-labeled mouse and human cells, and fragments of the uterine tube are engrafted by using minimally traumatic dorsal incision surgery. Transplanted cells and their outgrowths are easily located in the ovarian fat pad for over 40 days. Long-term transplantation of the entire uterine tube allows correct preservation of all principle tissue components, and does not result in adverse side effects, such as fibrosis and inflammation. Our approach should be uniquely applicable for answering important biological questions such as differentiation, regenerative and neoplastic potential of specific cell populations. Furthermore, it should be suitable for studies of microenvironmental factors in normal development and cancer.

We describe an ovarian fad pad transplantation assay suitable for studies of normal and transformed epithelia of the female reproductive tract. The mouse fat pad allows transplantation of large tissue fragments, is easily accessible for surgery and imaging, and provides the most favorable native environment for tissues of M&#252;llerian origin.

Orthotopic transplantation assays in mice are invaluable for studies of cell regeneration and neoplastic transformation. Common approaches for orthotopic ...

Laparoscopic Radical Left Pancreatectomy for Pancreatic Cancer: Surgical Strategy and Technique Video

Radical resection margins, resection of Gerota&#8217;s (perirenal) fascia, and adequate lymph node dissection are crucial for an adequate oncological resection of left-sided pancreatic cancer. Several surgical techniques have been described in recent years, but few were specifically designed for minimally invasive approaches. This study describes and demonstrates a standardized and reproducible technique for an adequate oncological resection of pancreatic cancer: laparoscopic radical left pancreatectomy (LRLP).
A 61-year-old woman presented with an incidental finding of a 3 cm mass in the left pancreas suspect for malignancy. Imaging did not reveal distant metastases, central vascular involvement, or morbid obesity, hence the patient was suitable for LRLP. This study describes the main steps of LRLP for pancreatic cancer. First, the lesser sac is opened by transecting the gastrocolic ligament. The splenic flexure of the colon is mobilized and the inferior border of the pancreas including Gerota&#39;s fascia is dissected down to the inferior border of the spleen. The pancreas is tunneled and hung, including Gerota&#8217;s fascia with a vessel loop. At the pancreatic neck, a tunnel is created between the pancreas and the portal vein, likewise a vessel loop is passed. The pancreas is then transected using the graded compression technique with an endostapler. Both the splenic vein and artery are transected before completing the resection. The entire specimen is extracted in a retrieval bag via a small Pfannenstiel incision.
Duration of the surgery was 210 min with 250 mL blood loss. Pathology revealed a R0-resection (&#62;1 mm) of a well-to-moderately differentiated adenocarcinoma originating from an intraductal papillary mucinous neoplasm. A total of 15 tumor-negative lymph nodes were resected. This is a detailed description of LRLP for left-sided pancreatic cancer as is currently being used within the international, multicenter randomized DIPLOMA (Distal Pancreatectomy Minimally Invasive or Open for PDAC) trial.

Oncologically safe left pancreatectomy requires radical resection (R0), Gerota&#8217;s (perirenal) fascia resection, and adequate lymph node dissection. This study describes the technical details of laparoscopic radical left pancreatectomy (LRLP), used in the first international multicenter randomized trial comparing minimally invasive with open left pancreatectomy for pancreatic cancer, the DIPLOMA trial.

Radical resection margins, resection of Gerota&#8217;s (perirenal) fascia, and adequate lymph node dissection are crucial for an adequate oncological ...

Fertility Preservation in Patients with Severe Ovarian Dysfunction

Ovarian function progressively declines during aging and in some pathophysiological conditions including karyotype abnormality, autoimmune diseases, chemo- and radiation-therapies, as well as ovarian surgeries. In unmarried women with severe ovarian dysfunction, fertility preservation is important for future pregnancies. Although oocyte cryopreservation is an established method for fertility preservation, these patients could only preserve a limited number of oocytes even after ovarian hyperstimulation, leading to repeated stimulations to ensure sufficient oocytes to guarantee future pregnancy. To solve this issue, we have recently developed a drug-free in vitro activation (IVA) procedure, which enable us to stimulate early stages of ovarian follicles to develop to the preantral follicle stage. These preantral follicles can respond to the unique protocol of gonadotropin stimulation, resulting in increased number of retrieved oocytes per ovarian stimulation for cryopreservation. The drug-free IVA comprised from the surgical approach and ovarian stimulation. We removed a part of cortex from one or both ovaries from patients under laparoscopic surgery. The ovarian cortical tissues were cut into small cubes to disrupt the Hippo signaling pathway and stimulate the development of early stage follicles. These cubes were grafted orthotropically into remaining ovaries as well as beneath the serosa of both Fallopian tubes. We have already published the surgical procedure of the drug-free IVA and the protocol of subsequent ovarian stimulation, but herein we present the details of laboratory methods required for drug-free IVA.

We present details of laboratory procedures for drug-free in vitro activation (IVA) of ovarian follicles for patients with severe ovarian dysfunction. This method could increase the number of retrievable oocytes per ovarian hyperstimulation and benefit fertility preservation for those patients.

Ovarian function progressively declines during aging and in some pathophysiological conditions including karyotype abnormality, autoimmune diseases, ...