{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T12:51:19Z","timestamp":1782910279851,"version":"3.54.5"},"reference-count":62,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2019,11,30]],"date-time":"2019-11-30T00:00:00Z","timestamp":1575072000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005010","name":"Associazione Italiana per la Ricerca sul Cancro","doi-asserted-by":"publisher","award":["IG 2013-14272"],"award-info":[{"award-number":["IG 2013-14272"]}],"id":[{"id":"10.13039\/501100005010","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Background: Ovaries are sensitive to chemotherapy, which may lead to early depletion of primordial follicle reserve. One strategy for gonadal function preservation is temporary ovarian suppression with Gonadotropin Releasing Hormone agonists (GnRHa) during chemotherapy. To date, GnRHa protective mechanism of action remains not fully elucidated. Methods: We collected 260 immature cumulus cell-oocyte complexes (COC) from 111 women &lt; 38 years old, with a normal ovarian reserve. The COC were randomly assigned to the following groups: (a) control; culture with the addition of (b) GnRHa; (c) cyclophosphamide; (d) cyclophosphamide plus GnRHa. After in vitro treatments, RNA and proteins were extracted from oocytes and cumulus cells (CC), separately. Potential effects of drugs were evaluated on GnRH receptors, apoptosis pathways, ceramide pathway, and glutathione synthesis by quantitative PCR and, whenever possible, by Western blot. Results: Cyclophosphamide triggered activation of the extrinsic pathway of apoptosis mediated by BAX in CC. The co-administration of GnRHa inhibited the apoptosis pathway in CC. According to our model, the GnRHa does not directly act on oocytes, which do not express GnRH receptors. Moreover, glutathione synthesis was decreased after GnRHa treatment both in CC and oocytes. Conclusion: Our data suggest that the protective mechanisms induced by GnRHa is mediated by an anti-apoptotic effect on CC.<\/jats:p>","DOI":"10.3390\/ijms20236045","type":"journal-article","created":{"date-parts":[[2019,12,2]],"date-time":"2019-12-02T10:50:45Z","timestamp":1575283845000},"page":"6045","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Gonadotropin Releasing Hormone Agonists Have an Anti-apoptotic Effect on Cumulus Cells"],"prefix":"10.3390","volume":"20","author":[{"given":"Paola","family":"Scaruffi","sequence":"first","affiliation":[{"name":"UOS Physiopathology of Human Reproduction, IRCCS Ospedale Policlinico San Martino, 16132 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara","family":"Stigliani","sequence":"additional","affiliation":[{"name":"UOS Physiopathology of Human Reproduction, IRCCS Ospedale Policlinico San Martino, 16132 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Cardinali","sequence":"additional","affiliation":[{"name":"Breast Unit, IRCCS Ospedale Policlinico San Martino, 16132 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia","family":"Massarotti","sequence":"additional","affiliation":[{"name":"Department of Neurosciences, Rehabilitation, Ophthalmology, Genetics, Maternal and Child Health, University of Genoa, 16132 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1797-5296","authenticated-orcid":false,"given":"Matteo","family":"Lambertini","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, University of Genoa, 16132 Genoa, Italy"},{"name":"U.O.C. Clinica di Oncologia Medica, IRCCS Ospedale Policlinico San Martino, 16132 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fausta","family":"Sozzi","sequence":"additional","affiliation":[{"name":"UOS Physiopathology of Human Reproduction, IRCCS Ospedale Policlinico San Martino, 16132 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chiara","family":"Dellepiane","sequence":"additional","affiliation":[{"name":"Breast Unit, IRCCS Ospedale Policlinico San Martino, 16132 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Domenico Franco","family":"Merlo","sequence":"additional","affiliation":[{"name":"Infrastruttura Ricerca e Statistica, Azienda Unit\u00e0 Sanitaria Locale\u2014IRCCS di Reggio Emilia, 42123 Reggio Emilia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2546-2626","authenticated-orcid":false,"given":"Paola","family":"Anserini","sequence":"additional","affiliation":[{"name":"UOS Physiopathology of Human Reproduction, IRCCS Ospedale Policlinico San Martino, 16132 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lucia","family":"Del Mastro","sequence":"additional","affiliation":[{"name":"Breast Unit, IRCCS Ospedale Policlinico San Martino, 16132 Genoa, Italy"},{"name":"Department of Internal Medicine, University of Genoa, 16132 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,11,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Wallace, W.H., and Kelsey, T.W. (2010). Human ovarian reserve from conception to the menopause. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0008772"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1093\/humupd\/7.6.535","article-title":"The effects of radiotherapy and chemotherapy on female reproduction","volume":"7","author":"Meirow","year":"2001","journal-title":"Hum. Reprod. Update"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"10159","DOI":"10.1158\/0008-5472.CAN-07-2042","article-title":"A novel ovarian xenografting model to characterize the impact of chemotherapy agents on human primordial follicle reserve","volume":"67","author":"Oktem","year":"2007","journal-title":"Cancer Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"525","DOI":"10.1093\/humupd\/dms022","article-title":"How do chemotherapeutic agents damage the ovary?","volume":"18","author":"Morgan","year":"2012","journal-title":"Hum. Reprod. Update"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1657","DOI":"10.1056\/NEJMra1614676","article-title":"Fertility Preservation in Women","volume":"377","author":"Donnez","year":"2017","journal-title":"N. Engl. J. Med."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Lambertini, M., Del Mastro, L., Pescio, M.C., Andersen, C.Y., Azim, H.A., Peccatori, F.A., Costa, M., Revelli, A., Salvagno, F., and Gennari, A. (2016). Cancer and fertility preservation: International recommendations from an expert meeting. BMC Med., 14.","DOI":"10.1186\/s12916-015-0545-7"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.ejca.2016.10.034","article-title":"Temporary ovarian suppression during chemotherapy to preserve ovarian function and fertility in breast cancer patients: A GRADE approach for evidence evaluation and recommendations by the Italian Association of Medical Oncology","volume":"71","author":"Lambertini","year":"2017","journal-title":"Eur. J. Cancer"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1994","DOI":"10.1200\/JCO.2018.78.1914","article-title":"Fertility preservation in patients with cancer: ASCO clinical practice guideline update","volume":"36","author":"Oktay","year":"2018","journal-title":"J. Clin. Oncol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1016\/j.breast.2017.07.017","article-title":"ESO-ESMO 3rd international consensus guidelines for breast cancer in young women (BCY3)","volume":"35","author":"Pagani","year":"2017","journal-title":"Breast"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/j.ctrv.2018.11.006","article-title":"Ovarian protection with gonadotropin-releasing hormone agonists during chemotherapy in cancer patients: From biological evidence to clinical application","volume":"72","author":"Lambertini","year":"2019","journal-title":"Cancer Treat. Rev."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1981","DOI":"10.1200\/JCO.2018.78.0858","article-title":"Gonadotropin-releasing hormone agonists during chemotherapy for preservation of ovarian function and fertility in premenopausal patients with early breast cancer: A systematic review and meta-analysis of individual patient-level data","volume":"36","author":"Lambertini","year":"2018","journal-title":"J. Clin. Oncol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2568","DOI":"10.1200\/JCO.2015.65.8864","article-title":"No Evidence for the Benefit of Gonadotropin-Releasing Hormone Agonist in Preserving Ovarian Function and Fertility in Lymphoma Survivors Treated With Chemotherapy: Final Long-Term Report of a Prospective Randomized Trial","volume":"34","author":"Demeestere","year":"2016","journal-title":"J. Clin. Oncol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1002\/uog.18934","article-title":"Gonadotropin-releasing hormone agonists for ovarian protection during cancer chemotherapy: Systematic review and meta-analysis","volume":"51","author":"Senra","year":"2018","journal-title":"Ultrasound Obstet. Gynecol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2408","DOI":"10.1093\/annonc\/mdv374","article-title":"Ovarian suppression using luteinizing hormone-releasing hormone agonists during chemotherapy to preserve ovarian function and fertility of breast cancer patients: A meta-analysis of randomized studies","volume":"26","author":"Lambertini","year":"2015","journal-title":"Ann. Oncol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1001\/jamaoncol.2015.3251","article-title":"Gonadotropin-releasing hormone agonists for ovarian function preservation in premenopausal women undergoing chemotherapy for early-stage breast cancer: A systematic review and meta-analysis","volume":"2","author":"Munhoz","year":"2016","journal-title":"JAMA Oncol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1159\/000454983","article-title":"Protecting effects of gonadotropin releasing hormone agonist on chemotherapy-induced ovarian damage in premenopausal breast cancer patients: A systematic review and meta-analysis","volume":"12","author":"Bai","year":"2017","journal-title":"Breast Care"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1333","DOI":"10.1016\/j.fertnstert.2004.12.030","article-title":"Acute depletion of murine primordial follicle reserve by gonadotropin-releasing hormone antagonists","volume":"83","author":"Danforth","year":"2005","journal-title":"Fertil. Steril."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1007\/s00404-015-3831-x","article-title":"Effects of GnRHa on early embryonic development in mice receiving cyclophosphamide","volume":"293","author":"Kanter","year":"2016","journal-title":"Arch. Gynecol. Obstet."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1294","DOI":"10.1093\/humrep\/deh257","article-title":"The GnRH antagonist cetrorelix reduces cyclophosphamide-induced ovarian follicular destruction in mice","volume":"19","author":"Meirow","year":"2004","journal-title":"Hum. Reprod."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"25","DOI":"10.2739\/kurumemedj.54.25","article-title":"GnRH agonist acts as ovarian protection in chemotherapy induced gonadotoxicity: An experiment using a rat model","volume":"54","author":"Matsuo","year":"2007","journal-title":"Kurume Med. J."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1179\/1973947813Y.0000000069","article-title":"Goserelin can inhibit ovarian cancer proliferation and simultaneously protect ovarian function from cisplatin: An in vitro and in vivo study","volume":"25","author":"Zhang","year":"2013","journal-title":"J. Chemother."},{"key":"ref_22","first-page":"3651","article-title":"A luteinizing hormone-releasing hormone agonist for the prevention of chemotherapy-induced ovarian follicular loss in rats","volume":"45","author":"Ataya","year":"1985","journal-title":"Cancer Res."},{"key":"ref_23","first-page":"7252","article-title":"Inhibition of rat ovarian [3H]thymidine uptake by luteinizing hormone-releasing hormone agonists: A possible mechanism for preventing damage by cytotoxic agents","volume":"48","author":"Ataya","year":"1988","journal-title":"Cancer Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"861","DOI":"10.1038\/bjc.1990.192","article-title":"Protective effects of D-Trp6-luteinising hormone-releasing hormone microcapsules against cyclophosphamideinduced gonadotoxicity in female rats","volume":"61","author":"Bokser","year":"1990","journal-title":"Br. J. Cancer"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1186\/1477-7827-11-16","article-title":"Combination of a GnRH agonist with an antagonist prevents flare-up effects and protects primordial ovarian follicles in the rat ovary from cisplatin-induced toxicity: A controlled experimental animal study","volume":"11","author":"Li","year":"2013","journal-title":"Reprod. Biol. Endocrinol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Horicks, F., Van Den Steen, G., Houben, S., Englert, Y., and Demeestere, I. (2015). Folliculogenesis Is Not Fully Inhibited during GnRH Analogues Treatment in Mice Challenging Their Efficiency to Preserve the Ovarian Reserve during Chemotherapy in This Model. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0137164"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1093\/molehr\/gay005","article-title":"Both in vivo FSH depletion and follicular exposure to Gonadotrophin-releasing hormone analogues in vitro are not effective to prevent follicular depletion during chemotherapy in mice","volume":"24","author":"Horicks","year":"2018","journal-title":"Mol. Hum. Reprod."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1157:e1","DOI":"10.1016\/j.fertnstert.2013.12.028","article-title":"Goserelin fosters bone elongation but does not prevent ovarian damage in cyclophosphamide-treated prepubertal mice","volume":"101","author":"Detti","year":"2014","journal-title":"Fertil. Steril."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1089","DOI":"10.1093\/humrep\/dev037","article-title":"Gonadotrophin-releasing hormone agonists for fertility preservation: Unraveling the enigma?","volume":"30","author":"Hasky","year":"2015","journal-title":"Hum. Reprod."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1095\/biolreprod52.2.365","article-title":"Luteinizing hormone-releasing hormone agonist inhibits cyclophosphamide induced ovarian follicular depletion in rhesus monkeys","volume":"52","author":"Ataya","year":"1995","journal-title":"Biol. Reprod."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1159\/000096062","article-title":"Direct protection by a gonadotropin-releasing hormone analog from doxorubicin-induced granulosa cell damage","volume":"63","author":"Imai","year":"2007","journal-title":"Gynecol. Obstet. Investig."},{"key":"ref_32","first-page":"2912","article-title":"GnRH agonist leuprolide acetate does not confer any protection against ovarian damage induced by chemotherapy and radiation in vitro","volume":"30","author":"Bildik","year":"2015","journal-title":"Hum. Reprod."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"3877","DOI":"10.1210\/en.2013-1341","article-title":"Characterization of the gonadotropin releasing hormone receptor (GnRHR) expression and activity in the female mouse ovary","volume":"154","author":"Torrealday","year":"2013","journal-title":"Endocrinology"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2189","DOI":"10.1007\/s00018-004-4001-7","article-title":"Alternative splicing of Bcl-2-related genes: Functional comnseguences and potential therapeutic applications","volume":"61","author":"Akgul","year":"2004","journal-title":"Cell Mol. Life Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1093\/humupd\/dmn022","article-title":"GnRH-analogues and oral contraceptives for fertility preservation in women during chemotherapy","volume":"14","author":"Blumenfeld","year":"2008","journal-title":"Hum. Reprod. Update"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1044","DOI":"10.1634\/theoncologist.12-9-1044","article-title":"How to preserve fertility in young women exposed to chemotherapy? The role of GnRH agonist cotreatment in addition to cryopreservation of embrya, oocytes, or ovaries","volume":"12","author":"Blumenfeld","year":"2007","journal-title":"Oncologist"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1179558119864584","DOI":"10.1177\/1179558119864584","article-title":"Potential mechanisms of ovarian protection with gonadotropin-releasing hormone agonist in breast cancer patients: A review","volume":"13","author":"Poggio","year":"2019","journal-title":"Clin. Med. Insights Reprod. Health"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1609","DOI":"10.1016\/j.fertnstert.2009.02.054","article-title":"Prevention of paclitaxel and cisplatin induced ovarian damage in rats by a gonadotropin-releasing hormone agonist","volume":"93","author":"Ozcelik","year":"2010","journal-title":"Fertil. Steril."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1016\/j.ejogrb.2009.12.028","article-title":"Protective effect of a gonadotropin-releasing hormone analogue on chemotherapeutic agent-induced ovarian gonadotoxicity: A mouse model","volume":"149","author":"Tan","year":"2010","journal-title":"Eur. J. Obstet. Gynecol. Reprod. Biol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"894","DOI":"10.1631\/jzus.B1100369","article-title":"Influence of gonadotropin-releasing hormone agonist on the effect of chemotherapy upon ovarian cancer and the prevention of chemotherapy-induced ovarian damage: An experimental study with nu\/nu athymic mice","volume":"13","author":"Lin","year":"2012","journal-title":"J. Zhejiang Univ. Sci. B"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1023","DOI":"10.1007\/s00404-012-2658-y","article-title":"Effect of a gonadotropin-releasing hormone analogue on cyclophosphamide-induced ovarian toxicity in adult mice","volume":"287","author":"Kishk","year":"2013","journal-title":"Arch. Gynecol. Obstet."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1080\/01443615.2017.1306839","article-title":"A gonadotropin-releasing hormone agonist for the prevention of docetaxel-induced gonadal damage","volume":"37","author":"Park","year":"2017","journal-title":"J. Obstet. Gynaecol."},{"key":"ref_43","first-page":"3810","article-title":"Luteinizing hormone releasing hormone induced nuclear factor kappaB activation and inhibits apoptosis in ovarian cancer cells","volume":"85","author":"Grundker","year":"2000","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1006\/dbio.2000.9863","article-title":"Oocyte-granulosa cell heterologous gap junctions are required for the coordination of nuclear and cytoplasmic meiotic competence","volume":"226","author":"Carabatsos","year":"2000","journal-title":"Dev. Biol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1093\/humupd\/dmm040","article-title":"Oocyte-secreted factors: Regulators of cumulus cell function and oocyte quality","volume":"14","author":"Gilchrist","year":"2008","journal-title":"Hum. Reprod. Update"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"673","DOI":"10.1530\/jrf.0.0990673","article-title":"Oxygen tension in the oviduct and uterus of rhesus monkeys, hamsters and rabbits","volume":"99","author":"Fischer","year":"1993","journal-title":"J. Reprod. Fertil."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1093\/humrep\/15.suppl_2.199","article-title":"Toxic effects of oxygen on human embryo development","volume":"15","author":"Catt","year":"2000","journal-title":"Hum. Reprod."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1080\/01926230701320337","article-title":"Apoptosis: A Review of Programmed Cell Death","volume":"35","author":"Elmore","year":"2007","journal-title":"Toxicol. Pathol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1016\/S0092-8674(01)00237-9","article-title":"The TNF and TNF receptor superfamilies: Integrating mammalian biology","volume":"104","author":"Locksley","year":"2001","journal-title":"Cell"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"647","DOI":"10.1038\/nrc883","article-title":"The Bcl2 family: Regulators of the cellular life-or-death switch","volume":"2","author":"Cory","year":"2002","journal-title":"Nat. Rev. Cancer"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"4562","DOI":"10.1210\/jc.2006-1147","article-title":"Immunolocalization of gonadotropin-releasing hormone (GnRH)-I, GnRH-II, and type I GnRH receptor during follicular development in the human ovary","volume":"91","author":"Choi","year":"2006","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"3143","DOI":"10.1016\/j.bbagen.2012.09.008","article-title":"Glutathione synthesis","volume":"1830","author":"Lu","year":"2013","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_53","first-page":"303","article-title":"Role of glutathione in cancer pathophysiology and therapeutic interventions","volume":"9","author":"Singh","year":"2012","journal-title":"J. Exp. Ther. Oncol."},{"key":"ref_54","first-page":"3584","article-title":"Role of glutathione in the metabolism-dependent toxicity and chemotherapy of cyclophosphamide","volume":"41","author":"Gurtoo","year":"1981","journal-title":"Cancer Res."},{"key":"ref_55","first-page":"269","article-title":"Effect of the gonadotropin-releasing hormone analogue triptorelin on the occurrence of chemotherapy-induced early menopause in premenopausal women with breast cancer: A randomized trial","volume":"306","author":"Boni","year":"2011","journal-title":"JAMA"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"S649","DOI":"10.1016\/j.ajog.2018.11.1033","article-title":"Smart Diaphragm Study: Multi-omics profiling and cervical device measurements during pregnancy","volume":"220","author":"Liang","year":"2019","journal-title":"AJOG"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1002\/pmic.201600399","article-title":"Time, space, and disorder in the expanding proteome universe","volume":"17","author":"Minde","year":"2017","journal-title":"Proteomics"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1016\/j.ejogrb.2012.04.013","article-title":"Normal serum anti-M\u00fcllerian hormone levels in the general female population and the relationship with reproductive history","volume":"163","author":"Spada","year":"2012","journal-title":"Eur. J. Obstet. Gynecol. Reprod. Biol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"2519","DOI":"10.1093\/humrep\/dev232","article-title":"Storage time does not modify the gene expression profile of cryopreserved human metaphase II oocytes","volume":"30","author":"Stigliani","year":"2015","journal-title":"Hum. Reprod."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"i2","DOI":"10.1093\/humrep\/des200","article-title":"The oocyte","volume":"27","author":"Rienzi","year":"2012","journal-title":"Hum. Reprod."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1007\/s10495-012-0783-5","article-title":"The apoptotic transcriptome of the human MII oocyte: Characterization and age-related changes","volume":"18","author":"Santonocito","year":"2013","journal-title":"Apoptosis"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/23\/6045\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:38:56Z","timestamp":1760189936000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/23\/6045"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,11,30]]},"references-count":62,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2019,12]]}},"alternative-id":["ijms20236045"],"URL":"https:\/\/doi.org\/10.3390\/ijms20236045","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,11,30]]}}}