Name: op-ivm: combining in vitro maturation after oocyte retrieval with gynecological surgery
Uploaded: 2021-05-09T14:00:00
Description: Watch this Scientific Journal Video about OP-IVM: Combining In vitro Maturation after Oocyte Retrieval with Gynecological Surgery at JoVE.com

This work was supported by grants from the China National Key R&#38;D Program (no. 2017YFC1002000, 2018YFC1004001, 2019YFA0801400), the National Science Foundation of China (no. 81571386, 81730038), the CAMS Innovation Fund for Medical Sciences (2019-I2M-5-001), and the Special Research Project of Chinese Capital Health Development (2018-2-4095).

NOTE: Studies related to the OP-IVM method have been approved by the institutional review board (IRB) of Peking University Third Hospital and the Ethics Committee of Peking University (2014S2004). A summary of OP-IVM is shown in Figure 1. The step-by-step procedure will be introduced in the following section.
1. Introduction of OP-IVM to appropriate patients
<ol>
	<li>Identify potential patients who may benefit from OP-IVM such as those described in steps 1.1.1-...

<ol>
	<li>Ho, V. N. A., Braam, S. C., Pham, T. D., Mol, B. W., Vuong, L. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effectiveness+and+safety+of+in+vitro+maturation+of+oocytes+versus+in+vitro+fertilization+in+women+with+a+high+antral+follicle+count.">The effectiveness and safety of in vitro maturation of oocytes versus in vitro fertilization in women with a high antral follicle count.</a> Human Reproduction. 34 (6), 1055-1064 (2019).</li><li>Yang, Z. Y., Chian, R. 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S., 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=In+vitro+maturation+or+in+vitro+fertilization+for+women+with+polycystic+ovaries?+A+case-control+study+of+194+treatment+cycles.">In vitro maturation or in vitro fertilization for women with polycystic ovaries? A case-control study of 194 treatment cycles.</a> Fertility and Sterility. 98 (2), 355-360 (2012).</li><li>Ho, V. N. A., Braam, S. C., Pham, T. D., Mol, B. W., Vuong, L. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effectiveness+and+safety+of+in+vitro+maturation+of+oocytes+versus+in+vitro+fertilization+in+women+with+a+high+antral+follicle+count.">The effectiveness and safety of in vitro maturation of oocytes versus in vitro fertilization in women with a high antral follicle count.</a> Human Reproduction. 34 (6), 1055-1064 (2019).</li><li>Sahin, 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=Which+should+be+the+preferred+technique+during+laparoscopic+ovarian+cystectomy.">Which should be the preferred technique during laparoscopic ovarian cystectomy.</a> Reproductive Sciences. 24 (3), 393-399 (2017).</li><li>Xiao, 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=Impact+of+hemostatic+methods+on+ovarian+reserve+and+fertility+in+laparoscopic+ovarian+cystectomy.">Impact of hemostatic methods on ovarian reserve and fertility in laparoscopic ovarian cystectomy.</a> Experimental and Therapeutic. 17 (4), 2689-2693 (2019).</li><li>Song, X. -. 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=Enhancing+the+scope+of+in+vitro+maturation+for+fertility+preservation:+transvaginal+retrieval+of+immature+oocytes+during+endoscopic+gynaecological+procedures.">Enhancing the scope of in vitro maturation for fertility preservation: transvaginal retrieval of immature oocytes during endoscopic gynaecological procedures.</a> Human Reproduction. 35 (4), 837-846 (2020).</li><li>Magnusson, &. #. 1. 9. 7. ;., K&#228;llen, K., Thurin-Kjellberg, A., Bergh, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+number+of+oocytes+retrieved+during+IVF:+a+balance+between+efficacy+and+safety.">The number of oocytes retrieved during IVF: a balance between efficacy and safety.</a> Human Reproduction. 33 (1), 58-64 (2018).</li><li>Larose, H., et al., Wellik, D. 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=Gametogenesis:+A+journey+from+inception+to+conception.">Gametogenesis: A journey from inception to conception.</a> Current Topics in Developmental Biology. 132, 257-310 (2019).</li><li>Broer, S. L., Broekmans, F. J. M., Laven, J. S. E., Fauser, B. C. J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anti-M&#252;llerian+hormone:+ovarian+reserve+testing+and+its+potential+clinical+implications.">Anti-M&#252;llerian hormone: ovarian reserve testing and its potential clinical implications.</a> Human Reproduction Update. 20 (5), 688-701 (2014).</li><li>Arce, J. C., La Marca, A., Mirner Klein, B., Nyboe Andersen, A., Fleming, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antim&#252;llerian+hormone+in+gonadotropin+releasing-hormone+antagonist+cycles:+prediction+of+ovarian+response+and+cumulative+treatment+outcome+in+good-prognosis+patients.">Antim&#252;llerian hormone in gonadotropin releasing-hormone antagonist cycles: prediction of ovarian response and cumulative treatment outcome in good-prognosis patients.</a> Fertility and Sterility. 99 (6), 1644-1653 (2013).</li><li>Katayama, K. P., 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=Ultrasound-guided+transvaginal+needle+aspiration+of+follicles+for+in+vitro+fertilization.">Ultrasound-guided transvaginal needle aspiration of follicles for in vitro fertilization.</a> Obstetrics and Gynecology. 72 (2), 271-274 (1988).</li><li>Dellenbach, P., 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=Transvaginal+sonographically+controlled+follicle+puncture+for+oocyte+retrieval.">Transvaginal sonographically controlled follicle puncture for oocyte retrieval.</a> Fertility and Sterility. 44 (5), 656-662 (1985).</li><li>Hirata, T., 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=Concomitant+ovarian+drilling+and+oocyte+retrieval+by+laparoendoscopic+single-site+surgery+led+to+live+birth+using+in+vitro+maturation+of+oocyte+and+transfer+of+frozen-thawed+blastocyst+in+woman+with+polycystic+ovary+syndrome.">Concomitant ovarian drilling and oocyte retrieval by laparoendoscopic single-site surgery led to live birth using in vitro maturation of oocyte and transfer of frozen-thawed blastocyst in woman with polycystic ovary syndrome.</a> Journal of Obstetrics and Gynaecology Research. 40 (5), 1431-1435 (2014).</li><li>Deutinger, 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=Follicular+aspiration+for+in+vitro+fertilization:+sonographically+guided+transvaginal+versus+laparoscopic+approach.">Follicular aspiration for in vitro fertilization: sonographically guided transvaginal versus laparoscopic approach.</a> European Journal of Obstetrics, Gynecology, and Reproductive Biology. 26 (2), 127-133 (1987).</li><li>Tanbo, T., Henriksen, T., Magnus, O., Abyholm, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oocyte+retrieval+in+an+IVF+program.+A+comparison+of+laparoscopic+and+transvaginal+ultrasound-guided+follicular+puncture.">Oocyte retrieval in an IVF program. A comparison of laparoscopic and transvaginal ultrasound-guided follicular puncture.</a> Acta Obstetricia et Gynecologica Scandinavica. 67 (3), 243-246 (1988).</li><li>Nagy, Z. P., 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=Pregnancy+and+birth+after+intracytoplasmic+sperm+injection+of+in+vitro+matured+germinal-vesicle+stage+oocytes:+case+report.">Pregnancy and birth after intracytoplasmic sperm injection of in vitro matured germinal-vesicle stage oocytes: case report.</a> Fertility and Sterility. 65 (5), 1047-1050 (1996).</li><li>S&#246;derstr&#246;m-Anttila, V., M&#228;kinen, S., Tuuri, T., Suikkari, A. -. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Favourable+pregnancy+results+with+insemination+of+in+vitro+matured+oocytes+from+unstimulated+patients.">Favourable pregnancy results with insemination of in vitro matured oocytes from unstimulated patients.</a> Human Reproduction. 20 (6), 1534-1540 (2005).</li><li>Hwang, J. L., Lin, Y. H., Tsai, Y. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+maturation+and+fertilization+of+immature+oocytes:+a+comparative+study+of+fertilization+techniques.">In vitro maturation and fertilization of immature oocytes: a comparative study of fertilization techniques.</a> Journal of Assisted Reproduction and Genetics. 17 (1), 39-43 (2000).</li><li>Walls, M., Junk, S., Ryan, J. P., Hart, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IVF+versus+ICSI+for+the+fertilization+of+in-vitro+matured+human+oocytes.">IVF versus ICSI for the fertilization of in-vitro matured human oocytes.</a> Reproductive Biomedicine Online. 25 (6), 603-607 (2012).</li><li>Park, J. H., Jee, B. C., Kim, S. H. <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+normal+and+abnormal+fertilization+of+in+vitro-matured+human+oocyte+according+to+insemination+method.">Comparison of normal and abnormal fertilization of in vitro-matured human oocyte according to insemination method.</a> Journal of Obstetrics and Gynaecology Research. 42 (4), 417-421 (2016).</li><li>Ho, V. N. A., Pham, T. D., Le, A. H., Ho, T. M., Vuong, L. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Live+birth+rate+after+human+chorionic+gonadotropin+priming+in+vitro+maturation+in+women+with+polycystic+ovary+syndrome.">Live birth rate after human chorionic gonadotropin priming in vitro maturation in women with polycystic ovary syndrome.</a> Journal of Ovarian Research. 11 (1), 70 (2018).</li><li>Zheng, X., 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=Effect+of+hCG+priming+on+embryonic+development+of+immature+oocytes+collected+from+unstimulated+women+with+polycystic+ovarian+syndrome.">Effect of hCG priming on embryonic development of immature oocytes collected from unstimulated women with polycystic ovarian syndrome.</a> Reproductive Biology and Endocrinology. 10, 40 (2012).</li><li>Tannus, S., 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=Predictive+factors+for+live+birth+after+in+vitro+maturation+of+oocytes+in+women+with+polycystic+ovary+syndrome.">Predictive factors for live birth after in vitro maturation of oocytes in women with polycystic ovary syndrome.</a> Archives of Gynecology and Obstetrics. 297 (1), 199-204 (2018).</li><li>Cohen, Y., 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=Decreased+pregnancy+and+live+birth+rates+after+vitrification+of+in+vitro+matured+oocytes.">Decreased pregnancy and live birth rates after vitrification of in vitro matured oocytes.</a> Journal of Assisted Reproduction and Genetics. 35 (9), 1683-1689 (2018).</li><li>Yazdanpanah, F., Khalili, M. 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The OP-IVM method described in this article extends to conventional IVM applications and combines IVM after oocyte retrieval with routine gynecological surgery. Oocytes that would have been lost in the gynecological surgery can now be used for IVF-ET or FP without additional surgical risks. OP-IVM was first used to retrieve oocytes before ovarian drilling in PCOS patients. Its application soon expanded to infertile patients who need benign gynecological surgery and cancer or hematological disease patients who need chemor...

An erratum was issued for:&#160;<a href="https://www.jove.com/t/61647">OP-IVM: Combining In vitro Maturation after Oocyte Retrieval with Gynecological Surgery</a>. The Protocol and Representative Results sections were&#160;updated.
Step 3.2 of the Protocol was updated from:
Add 0.5 mL of IVM&#160;medium supplemented with 0.075 IU/mL of follicle-stimulating hormone (FSH) and 0.075 IU/mL of luteinizing hormone (LH) to each well of a 4-well plate. Cover the medium with oil.
to:
Add 0.5 mL of IVM&#160;medium supplemented with 0.75 IU/mL of follicle-stimulating hormone (FSH) and 0.75 IU/mL of luteinizing hormone (LH) to each well of a 4-well plate. Cover the medium with oil.
The legend for Figure 2 was updated from:
Figure 2:&#160;Morphological characteristics of representative images of oocytes. (A) COCs, (B) MII, (C) MI, (D) GV stage. Abbreviations: COCs = cumulus-oocyte complexes; GV = germinal vesicle; MI = metaphase I; MII = metaphase II.
to:
Figure 2:&#160;Morphological characteristics of representative images of oocytes. (A) COCs, (B) GV, (C) MI, (D) MII&#160;stage. Abbreviations: COCs = cumulus-oocyte complexes; GV = germinal vesicle; MI = metaphase I; MII = metaphase II.

An erratum was issued for:&#160;<a href="https://www.jove.com/t/61647">OP-IVM: Combining In vitro Maturation after Oocyte Retrieval with Gynecological Surgery</a>. The Protocol and Representative Results sections were&#160;updated.

Erratum: OP-IVM: Combining In vitro Maturation after Oocyte Retrieval with Gynecological Surgery

IVM is an ART in which human immature oocytes are cultured in vitro to maturation for IVF-ET or FP. In IVM, ovulation induction medications are not used, thus reducing pain, financial burden, and complications such as ovarian hyperstimulation syndrome (OHSS)1,2. In addition, IVM is particularly suitable for the FP of cancer patients and the infertility treatment of hormone-sensitive patients who may be unable to or have no time to receive ovulation induction therapy3. Therefore, although the number of oocytes retrieved, clinical pregnancy rate (CPR), and live birth rate (LBR) are lower than those of IVF4,5, IVM has its own unique advantages.
Infertile patients with endometrial lesions, hydrosalpinx, or ovarian cysts usually have gynecological surgery before ART treatment, and their oocytes are usually immature. OP-IVM uses guided transvaginal ultrasound to retrieve the immature oocytes and grow them in vitro until maturation for IVF-ET or FP. OP-IVM combines IVM after oocyte retrieval and gynecological surgery, thereby reducing complications that are common in&#160;controlled ovarian hyperstimulation cycles and saving time and money. For fertile patients, OP-IVM could serve as a &#34;fertility insurance&#34; while undergoing routine gynecological surgery.
Furthermore, damages caused by gynecological surgeries, such as electrocautery6,7 and ovarian tumor resection, could be reduced through oocyte retrieval before gynecological surgery. Therefore, compared with routine gynecological surgery, OP-IVM could reduce the number of operations during infertility treatment and prevent the loss of functional oocytes during ovarian surgery.
A previous study has shown that the additional procedure of oocyte retrieval would neither increase surgical complications and adverse pregnancy outcomes, nor prolong the hospital stay8. Some patients have given live birth through OP-IVM8, indicating the feasibility of this method. This paper describes characteristics of patients who may benefit from OP-IVM as well as procedures and critical points of OP-IVM and discusses the evaluation of human oocyte maturity.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>19 G single-lumen aspiration needles</td>
			<td>Cook, Australia</td>
			<td>K-OPS-7035-REH-ET</td>
			<td></td>
		</tr>
		<tr>
			<td>4-well plate</td>
			<td>Corning</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>70 &#956;m nylon cell strainer</td>
			<td>Falcon, USA</td>
			<td>352350</td>
			<td></td>
		</tr>
		<tr>
			<td>CO2 Incubator</td>
			<td>Thermo</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Culture oil</td>
			<td>Vitrolife, Sweden</td>
			<td>10029,OVOIL&#160;</td>
			<td>Step 3.2.</td>
		</tr>
		<tr>
			<td>FSH &#38; LH</td>
			<td>Ferring Reproductive Health, Germany</td>
			<td>MENOPUR&#174;</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass Pasteur pipette</td>
			<td>Hilgenberg GmbH, Germany</td>
			<td>3154102-26</td>
			<td></td>
		</tr>
		<tr>
			<td>G-MOPS medium</td>
			<td></td>
			<td></td>
			<td>pH-stable handling medium for washing the needle before puncturing</td>
		</tr>
		<tr>
			<td>IVM medium</td>
			<td>Origio, Denmark</td>
			<td>ART-1600-B</td>
			<td></td>
		</tr>
		<tr>
			<td>Laminar Flow Clean Benches</td>
			<td>ESCO</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Petri dish</td>
			<td>Thermo Fisher Scientific, Denmark</td>
			<td>263991</td>
			<td></td>
		</tr>
		<tr>
			<td>pH stable handing media designed to support the handling and manipulation of oocytes and embryos outside the incubator</td>
			<td>Vitrolife, Sweden</td>
			<td>10130, G-MOPS PLUS</td>
			<td>Step 7.1.</td>
		</tr>
		<tr>
			<td>Rinse solution</td>
			<td>Cook, Australia</td>
			<td>K-SIFB-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereoscope</td>
			<td>Nikon</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

op-ivm: combining in vitro maturation after oocyte retrieval with gynecological surgery

The use of in vitro maturation (IVM) before gynecological operation (OP-IVM) is an extension of conventional IVM that combines IVM following oocyte retrieval with routine gynecological surgery. OP-IVM is suitable for patients undergoing benign gynecological surgery who have the need for fertility preservation (FP) or infertility treatments such as in vitro fertilization and embryo transfer (IVF-ET). In the operating room, patients undergoing benign gynecological surgery are first anesthetized and receive ultrasound-guided immature follicle aspiration (IMFA) treatment. As the subsequent gynecological surgery is performed, the cumulus-oocyte complexes (COCs) are examined, and the immature COCs are transferred into the IVM medium and cultured for 28-32 hours in the IVF laboratory. After assessment, mature oocytes in the MII stage will be selected and cryopreserved in liquid nitrogen for FP or fertilized by intracytoplasmic sperm injection (ICSI) for IVF-ET. By combining IVM with gynecological surgery, immature oocytes that would have been discarded can be saved and used for assisted reproductive technology (ART). The procedure, significance and critical aspects of OP-IVM are described in this article.

Until December 2019, OP-IVM was used for fertility preservation of 274 patients. Embryological and reproductive outcomes of 158 patients between 2014 to 2016 were published in a previous paper8. The following example discusses the procedure followed for a PCOS patient receiving OP-IVM in 2016. The patient is a 28-year-old diagnosed with primary infertility, left adnexal cyst, and PCOS. She received laparoscopic cystectomy and OP-IVM on September 28th, 20...

Watch this Scientific Journal Video about OP-IVM: Combining In vitro Maturation after Oocyte Retrieval with Gynecological Surgery at JoVE.com

OP-IVM: Combining In vitro Maturation after Oocyte Retrieval with Gynecological Surgery

In vitro maturation (IVM) before gynecological operation (OP-IVM) combines IVM following oocyte retrieval with routine gynecological surgery and serves as an extension of conventional IVM applications for fertility preservation.

The use of in vitro maturation (IVM) before gynecological operation (OP-IVM) is an extension of conventional IVM that combines IVM following oocyte ...

The technology, OP-IVM, is an extension of conventional IVM protocol that combines IVM following oocyte retrieval with routine gynecological surgery. It is suitable for infertile or subfertile women who need to receive gynecological surgery before ART treatment or fertility preservation, polycystic ovary syndrome patients with clomiphene resistance who need a laparoscopic ovarian drilling surgery, infertile patients who are in need of benign gynecologic surgeries such as hysteroscopic myomectomy, polypectomy, transthoracic resection of septum, and the laparoscopic tube surgery, and the oophorocystectomy before ART treatment, and also patients with cancer and hematologic disease who are receiving chemoradiotherapy or radiotherapy, they can do this surgery for fertility preservation. Place the ultrasound probe inside the vagina to scan and record the number of follicles in both ovaries.Find the location closest to the ovary as the puncture site, avoiding the intestine, bladder, and large blood vessels. Wash the 19 gauge single lumen aspiration needle with a pH stable handling medium, then inject the needle into the ovaries under the guidance of ultrasound. Aspirate follicular fluid with the needle under a pressure of 80 to 90 millimeters of mercury, rotating the needle slightly to aspirate as much follicular fluid as possible.Use heparin to reduce follicular fluid viscosity during the aspiration. Adjust the probe's position to keep it as close as possible to the ovaries at all times and press the vaginal fornix with the proper force to reduce injury and bleeding. Make sure to complete follicular fluid aspiration within 25 to 30 minutes.Pull out the needle after finishing in one ovary, wash the needle with the handling medium, and puncture the other side using the same method. Working in a 37 degrees Celsius homothermal flat, prepare the handling media, Petri dish, and cell strainer. Rinse the culture tube and strainer with pre-warmed pH stable handling media.Filter the aspirated follicle fluid with a 70 micrometer nylon cell strainer, repeatedly rinsing the culture tube and strainer with pre-warmed pH stable handling medium. Ensure that all immature COCs are completely transferred to the culture dish. Transfer the blood into clean handling media.Then, transfer the cell strainer to a new dish and wash it. Aspirate and transfer the handling media with the COCs, granulosa cells and tissues to a new dish. Examine the COCs under the stereoscope with 40 times magnification.Quickly transfer the immatures into a pre-warmed IVM oocyte medium. Next, examine the COCs in the blood clot and in the cell strainer. Transfer the COCs and handling media into pre-warmed IVM oocyte medium.Culture the immature COCs for 28 to 32 hours at 37 degrees Celsius in humidified air containing 5%carbon dioxide and 5%oxygen. By December 2019, OP-IVM had been used for fertility preservation of 274 patients. One example was a 28 year old patient who was diagnosed with primary infertility, left adnexal cyst, and PCOS.She received laparoscopic cystectomy and OP-IVM in September 2016 and 27 immature COCs were obtained. After 28 hours in culture, seven oocytes in the MII stage were selected and fertilized by ICSI. After five days, two blastocysts were frozen.One was thawed in February 2017 and transferred into the uterus. The patient delivered a healthy girl in November. OP-IVM was first applied to the oocyte retrieval of infertile women with PCOS who are undergoing ovarian drilling operation, and now its applications can be expanded to the subfertile women who need to receive benign gynecological operation, even cancer women for fertility preservation.It indicates that the application range of IVM can be expanded by combining it with other traditional gynecological operations for female fertility preservation.

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A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the ...

Combining Augmented Reality and 3D Printing to Display Patient Models on a Smartphone

Augmented reality (AR) has great potential in education, training, and surgical guidance in the medical field. Its combination with three-dimensional (3D) ...

In vitro Assessment of Cardiac Reprogramming by Measuring Cardiac Specific Calcium Flux with a GCaMP3 Reporter

Cardiac reprogramming has become a potentially promising therapy to repair a damaged heart. By introducing multiple transcription factors, including ...

Fertility Sparing Procedure using Carbon Dioxide Fiber Laser Vaporization of Ovarian Endometrioma

The surgical management of endometrioma is still a matter of debate. Cystectomy, which is recognized as the standard technique, seems to be associated ...

Laparoscopic Oocyte Retrieval and Cryopreservation during Vaginoplasty for Treatment of Mayer-Rokitansky-Kuster-Hauser Syndrome

In Mayer-Rokitansky-Kuster-Hauser Syndrome (MRKHS) patients who are scheduled for laparoscopic vaginoplasty and have a desire for biological motherhood, ...

Lung Rapid Recovery Procurement Combined with Abdominal Normothermic Regional Perfusion in Controlled Donation after Circulatory Death

Controlled donation after circulatory death (cDCD) has contributed to increasing donor numbers all over the world. Experiences published in the last years ...

A Simple Pit Assay Protocol to Visualize and Quantify Osteoclastic Resorption In Vitro

A Simple Pit Assay Protocol to Visualize and Quantify Osteoclastic Resorption In Vitro

Mature osteoclasts are multinucleated cells that can degrade bone through the secretion of acids and enzymes. They play a crucial role in various diseases ...