Phytoestrogen exposure alters endometrial stromal cells and interferes with decidualization signaling
To investigate whether phytoestrogens (genistein and daidzein) alter in vitro decidualization of human endometrial stromal cells (ESCs). Isolated primary ESCs were exposed to phytoestrogens and decidualized in vitro. Academic fertility center. Twenty fertile oocyte donors attending the IVI Valencia...
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description | To investigate whether phytoestrogens (genistein and daidzein) alter in vitro decidualization of human endometrial stromal cells (ESCs).
Isolated primary ESCs were exposed to phytoestrogens and decidualized in vitro.
Academic fertility center.
Twenty fertile oocyte donors attending the IVI Valencia clinic.
Treatment of ESC with phytoestrogens at 0, 10, 20, 50, and 100 μM.
The ESC proliferation was analyzed by MTS assay. In vitro decidualization was induced in the presence of phytoestrogens by medroxyprogesterone acetate/cyclic adenosine 3′:5′ monophosphate and evaluated by prolactin (PRL) ELISA and F-actin immunostaining. The Ki67 proliferative marker was analyzed by immunofluorescence. The ESC apoptosis was assessed by annexin V/propidium iodide detection using flow cytometry. Estrogen (ERβ) and P receptor (PR) localization were evaluated by immunofluorescence.
The ESC exposed to 0, 19, 20, 50, and 100 μM of genistein, daidzein, and genistein + daidzein showed a dose-dependent proliferation decrease. After 48–96 hours of culture, this reduction was significant in the presence of 50 μM of phytoestrogens versus 10 μM untreated ESC. The ESC decidualized in the presence of phytoestrogens did not rearrange their cytoskeletons and showed a significant decrease in PRL secretion compared with untreated decidualized ESCs (dESCs). However, phytoestrogens did not alter proliferative status or the percentage of viable/apoptotic cells in dESC compared with untreated dESC. During decidualization, phytoestrogens induced the same nuclear translocation of ERβ and PR as the control dESC.
This study reveals that high doses of phytoestrogens could affect the in vitro decidualization process.
La exposición a fitoestrógenos altera células del estroma endometrial e interfiere con la señalización de la decidualización
Investigar si los fitoestrógenos (genisteína y daidzeína) alteran la decidualización de las células del estroma endometrial humano (CEE) in vitro.
Células estromales endometriales (CEE) primarias aisladas fueron expuestas a fitoestrógenos y decidualizadas in vitro.
Centro académico de fertilidad.
Veinte donantes de ovocitos fértiles que han acudido a la clínica IVI Valencia.
Tratamiento de las CEE con fitoestrógenos a 0, 10, 20, 50 y 100 μM.
La proliferación de CEE se analizó mediante el test MTS. La decidualización in vitro fue inducida en presencia de fitoestrógenos con acetato de medroxiprogesterona / adenosina cíclica monofosfato 3´:5´y evaluado por prolactina |
doi_str_mv | 10.1016/j.fertnstert.2019.06.014 |
format | Article |
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Isolated primary ESCs were exposed to phytoestrogens and decidualized in vitro.
Academic fertility center.
Twenty fertile oocyte donors attending the IVI Valencia clinic.
Treatment of ESC with phytoestrogens at 0, 10, 20, 50, and 100 μM.
The ESC proliferation was analyzed by MTS assay. In vitro decidualization was induced in the presence of phytoestrogens by medroxyprogesterone acetate/cyclic adenosine 3′:5′ monophosphate and evaluated by prolactin (PRL) ELISA and F-actin immunostaining. The Ki67 proliferative marker was analyzed by immunofluorescence. The ESC apoptosis was assessed by annexin V/propidium iodide detection using flow cytometry. Estrogen (ERβ) and P receptor (PR) localization were evaluated by immunofluorescence.
The ESC exposed to 0, 19, 20, 50, and 100 μM of genistein, daidzein, and genistein + daidzein showed a dose-dependent proliferation decrease. After 48–96 hours of culture, this reduction was significant in the presence of 50 μM of phytoestrogens versus 10 μM untreated ESC. The ESC decidualized in the presence of phytoestrogens did not rearrange their cytoskeletons and showed a significant decrease in PRL secretion compared with untreated decidualized ESCs (dESCs). However, phytoestrogens did not alter proliferative status or the percentage of viable/apoptotic cells in dESC compared with untreated dESC. During decidualization, phytoestrogens induced the same nuclear translocation of ERβ and PR as the control dESC.
This study reveals that high doses of phytoestrogens could affect the in vitro decidualization process.
La exposición a fitoestrógenos altera células del estroma endometrial e interfiere con la señalización de la decidualización
Investigar si los fitoestrógenos (genisteína y daidzeína) alteran la decidualización de las células del estroma endometrial humano (CEE) in vitro.
Células estromales endometriales (CEE) primarias aisladas fueron expuestas a fitoestrógenos y decidualizadas in vitro.
Centro académico de fertilidad.
Veinte donantes de ovocitos fértiles que han acudido a la clínica IVI Valencia.
Tratamiento de las CEE con fitoestrógenos a 0, 10, 20, 50 y 100 μM.
La proliferación de CEE se analizó mediante el test MTS. La decidualización in vitro fue inducida en presencia de fitoestrógenos con acetato de medroxiprogesterona / adenosina cíclica monofosfato 3´:5´y evaluado por prolactina (PRL)-ELISA de prolactina (PRL) e inmunotinción actina F. El marcador proliferativo Ki67 fue analizado por inmunofluorescencia. La apoptosis de las CEE fue evaluada mediante la detección de anexina V / yoduro de propidio mediante citometría de flujo. La localización de los receptores de estrógeno (ERβ) y del receptor P de progesterona (PR) fue evaluado por inmunofluorescencia.
Las CEE expuestas a 0, 19, 20, 50 y 100 mM de genisteína, daidzeína y genisteína y daidzeína mostraron una disminución de la proliferación dosis dependiente. Después de 48-96 horas de cultivo, esta reducción fue significativa en presencia de 50 mM de fitoestrógenos versus 10 μM de CEE sin tratar. Las CEE decidualizadas en presencia de los fitoestrógenos no reorganizaron sus citoesqueletos y mostraron una disminución significativa en la secreción de PRL en comparación con las CEE decidualizados (dCEE) no tratadas. Sin embargo, los fitoestrógenos no alteraron el estado proliferativo o el porcentaje de células viables / apoptóticas en dCEE en comparación con dCEE no tratadas. Durante la decidualización, los fitoestrógenos indujeron la misma translocación nuclear de ERβ y PR que en las dCEE control.
Este estudio revela que altas dosis de fitoestrógenos podrían afectar el proceso de decidualización in vitro.</description><identifier>ISSN: 0015-0282</identifier><identifier>EISSN: 1556-5653</identifier><identifier>DOI: 10.1016/j.fertnstert.2019.06.014</identifier><identifier>PMID: 31371049</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Cell Proliferation - drug effects ; Cell Proliferation - physiology ; Cells, Cultured ; Decidua - cytology ; Decidua - drug effects ; Decidua - physiology ; decidualization ; Dose-Response Relationship, Drug ; endocrine disruptors ; endometrium ; Endometrium - cytology ; Endometrium - drug effects ; Endometrium - physiology ; Female ; Genistein - pharmacology ; Humans ; Isoflavones - pharmacology ; Phytoestrogens ; Phytoestrogens - pharmacology ; Signal Transduction - drug effects ; Signal Transduction - physiology ; steroid receptors ; Stromal Cells - drug effects ; Stromal Cells - physiology</subject><ispartof>Fertility and sterility, 2019-11, Vol.112 (5), p.947-958.e3</ispartof><rights>2019 The Authors</rights><rights>Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c424t-75ad4eef31e12c5a8eb833537c74a48a7084dc826d3e4e70dcd73fab6fe06e303</citedby><cites>FETCH-LOGICAL-c424t-75ad4eef31e12c5a8eb833537c74a48a7084dc826d3e4e70dcd73fab6fe06e303</cites><orcidid>0000-0002-7620-8081</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.fertnstert.2019.06.014$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>314,778,782,3539,27907,27908,45978</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31371049$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Salsano, Stefania</creatorcontrib><creatorcontrib>Pérez-Debén, Silvia</creatorcontrib><creatorcontrib>Quiñonero, Alicia</creatorcontrib><creatorcontrib>González-Martín, Roberto</creatorcontrib><creatorcontrib>Domínguez, Francisco</creatorcontrib><title>Phytoestrogen exposure alters endometrial stromal cells and interferes with decidualization signaling</title><title>Fertility and sterility</title><addtitle>Fertil Steril</addtitle><description>To investigate whether phytoestrogens (genistein and daidzein) alter in vitro decidualization of human endometrial stromal cells (ESCs).
Isolated primary ESCs were exposed to phytoestrogens and decidualized in vitro.
Academic fertility center.
Twenty fertile oocyte donors attending the IVI Valencia clinic.
Treatment of ESC with phytoestrogens at 0, 10, 20, 50, and 100 μM.
The ESC proliferation was analyzed by MTS assay. In vitro decidualization was induced in the presence of phytoestrogens by medroxyprogesterone acetate/cyclic adenosine 3′:5′ monophosphate and evaluated by prolactin (PRL) ELISA and F-actin immunostaining. The Ki67 proliferative marker was analyzed by immunofluorescence. The ESC apoptosis was assessed by annexin V/propidium iodide detection using flow cytometry. Estrogen (ERβ) and P receptor (PR) localization were evaluated by immunofluorescence.
The ESC exposed to 0, 19, 20, 50, and 100 μM of genistein, daidzein, and genistein + daidzein showed a dose-dependent proliferation decrease. After 48–96 hours of culture, this reduction was significant in the presence of 50 μM of phytoestrogens versus 10 μM untreated ESC. The ESC decidualized in the presence of phytoestrogens did not rearrange their cytoskeletons and showed a significant decrease in PRL secretion compared with untreated decidualized ESCs (dESCs). However, phytoestrogens did not alter proliferative status or the percentage of viable/apoptotic cells in dESC compared with untreated dESC. During decidualization, phytoestrogens induced the same nuclear translocation of ERβ and PR as the control dESC.
This study reveals that high doses of phytoestrogens could affect the in vitro decidualization process.
La exposición a fitoestrógenos altera células del estroma endometrial e interfiere con la señalización de la decidualización
Investigar si los fitoestrógenos (genisteína y daidzeína) alteran la decidualización de las células del estroma endometrial humano (CEE) in vitro.
Células estromales endometriales (CEE) primarias aisladas fueron expuestas a fitoestrógenos y decidualizadas in vitro.
Centro académico de fertilidad.
Veinte donantes de ovocitos fértiles que han acudido a la clínica IVI Valencia.
Tratamiento de las CEE con fitoestrógenos a 0, 10, 20, 50 y 100 μM.
La proliferación de CEE se analizó mediante el test MTS. La decidualización in vitro fue inducida en presencia de fitoestrógenos con acetato de medroxiprogesterona / adenosina cíclica monofosfato 3´:5´y evaluado por prolactina (PRL)-ELISA de prolactina (PRL) e inmunotinción actina F. El marcador proliferativo Ki67 fue analizado por inmunofluorescencia. La apoptosis de las CEE fue evaluada mediante la detección de anexina V / yoduro de propidio mediante citometría de flujo. La localización de los receptores de estrógeno (ERβ) y del receptor P de progesterona (PR) fue evaluado por inmunofluorescencia.
Las CEE expuestas a 0, 19, 20, 50 y 100 mM de genisteína, daidzeína y genisteína y daidzeína mostraron una disminución de la proliferación dosis dependiente. Después de 48-96 horas de cultivo, esta reducción fue significativa en presencia de 50 mM de fitoestrógenos versus 10 μM de CEE sin tratar. Las CEE decidualizadas en presencia de los fitoestrógenos no reorganizaron sus citoesqueletos y mostraron una disminución significativa en la secreción de PRL en comparación con las CEE decidualizados (dCEE) no tratadas. Sin embargo, los fitoestrógenos no alteraron el estado proliferativo o el porcentaje de células viables / apoptóticas en dCEE en comparación con dCEE no tratadas. Durante la decidualización, los fitoestrógenos indujeron la misma translocación nuclear de ERβ y PR que en las dCEE control.
Este estudio revela que altas dosis de fitoestrógenos podrían afectar el proceso de decidualización in vitro.</description><subject>Cell Proliferation - drug effects</subject><subject>Cell Proliferation - physiology</subject><subject>Cells, Cultured</subject><subject>Decidua - cytology</subject><subject>Decidua - drug effects</subject><subject>Decidua - physiology</subject><subject>decidualization</subject><subject>Dose-Response Relationship, Drug</subject><subject>endocrine disruptors</subject><subject>endometrium</subject><subject>Endometrium - cytology</subject><subject>Endometrium - drug effects</subject><subject>Endometrium - physiology</subject><subject>Female</subject><subject>Genistein - pharmacology</subject><subject>Humans</subject><subject>Isoflavones - pharmacology</subject><subject>Phytoestrogens</subject><subject>Phytoestrogens - pharmacology</subject><subject>Signal Transduction - drug effects</subject><subject>Signal Transduction - physiology</subject><subject>steroid receptors</subject><subject>Stromal Cells - drug effects</subject><subject>Stromal Cells - physiology</subject><issn>0015-0282</issn><issn>1556-5653</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkEtuFDEQQC0EIkPgCshLNt0pf9rtLCEigBQJFrC2PHb1xKNue7DdkHAazsLJ8GgCLNlUqaRXv0cIZdAzYOpi30-Yayy1xZ4Du-xB9cDkI7Jhw6C6QQ3iMdkAsKEDrvkZeVbKHgAUG_lTciaYGBnIyw2ZPt3e14Sl5rTDSPHukMqakdq5zS4Uo08L1hzsTI_M0rLDeS7URv_rZ4iNaqdgod9DvaUeXfCrncMPW0OKtIRdbFXcPSdPJjsXfPGQz8mX67efr953Nx_ffbh6fdM5yWXtxsF6iTgJhoy7wWrcaiEGMbpRWqntCFp6p7nyAiWO4J0fxWS3akJQKECck1enuYecvq7tLbOEcjzYRkxrMZwrLUAr0A3VJ9TlVErGyRxyWGy-NwzM0bLZm3-WzdGyAWWa5db68mHLul3Q_238o7UBb04Atl-_BcymuIDRoQ8ZXTU-hf9v-Q2r65gZ</recordid><startdate>201911</startdate><enddate>201911</enddate><creator>Salsano, Stefania</creator><creator>Pérez-Debén, Silvia</creator><creator>Quiñonero, Alicia</creator><creator>González-Martín, Roberto</creator><creator>Domínguez, Francisco</creator><general>Elsevier Inc</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-7620-8081</orcidid></search><sort><creationdate>201911</creationdate><title>Phytoestrogen exposure alters endometrial stromal cells and interferes with decidualization signaling</title><author>Salsano, Stefania ; Pérez-Debén, Silvia ; Quiñonero, Alicia ; González-Martín, Roberto ; Domínguez, Francisco</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c424t-75ad4eef31e12c5a8eb833537c74a48a7084dc826d3e4e70dcd73fab6fe06e303</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Cell Proliferation - drug effects</topic><topic>Cell Proliferation - physiology</topic><topic>Cells, Cultured</topic><topic>Decidua - cytology</topic><topic>Decidua - drug effects</topic><topic>Decidua - physiology</topic><topic>decidualization</topic><topic>Dose-Response Relationship, Drug</topic><topic>endocrine disruptors</topic><topic>endometrium</topic><topic>Endometrium - cytology</topic><topic>Endometrium - drug effects</topic><topic>Endometrium - physiology</topic><topic>Female</topic><topic>Genistein - pharmacology</topic><topic>Humans</topic><topic>Isoflavones - pharmacology</topic><topic>Phytoestrogens</topic><topic>Phytoestrogens - pharmacology</topic><topic>Signal Transduction - drug effects</topic><topic>Signal Transduction - physiology</topic><topic>steroid receptors</topic><topic>Stromal Cells - drug effects</topic><topic>Stromal Cells - physiology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Salsano, Stefania</creatorcontrib><creatorcontrib>Pérez-Debén, Silvia</creatorcontrib><creatorcontrib>Quiñonero, Alicia</creatorcontrib><creatorcontrib>González-Martín, Roberto</creatorcontrib><creatorcontrib>Domínguez, Francisco</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Fertility and sterility</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Salsano, Stefania</au><au>Pérez-Debén, Silvia</au><au>Quiñonero, Alicia</au><au>González-Martín, Roberto</au><au>Domínguez, Francisco</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Phytoestrogen exposure alters endometrial stromal cells and interferes with decidualization signaling</atitle><jtitle>Fertility and sterility</jtitle><addtitle>Fertil Steril</addtitle><date>2019-11</date><risdate>2019</risdate><volume>112</volume><issue>5</issue><spage>947</spage><epage>958.e3</epage><pages>947-958.e3</pages><issn>0015-0282</issn><eissn>1556-5653</eissn><abstract>To investigate whether phytoestrogens (genistein and daidzein) alter in vitro decidualization of human endometrial stromal cells (ESCs).
Isolated primary ESCs were exposed to phytoestrogens and decidualized in vitro.
Academic fertility center.
Twenty fertile oocyte donors attending the IVI Valencia clinic.
Treatment of ESC with phytoestrogens at 0, 10, 20, 50, and 100 μM.
The ESC proliferation was analyzed by MTS assay. In vitro decidualization was induced in the presence of phytoestrogens by medroxyprogesterone acetate/cyclic adenosine 3′:5′ monophosphate and evaluated by prolactin (PRL) ELISA and F-actin immunostaining. The Ki67 proliferative marker was analyzed by immunofluorescence. The ESC apoptosis was assessed by annexin V/propidium iodide detection using flow cytometry. Estrogen (ERβ) and P receptor (PR) localization were evaluated by immunofluorescence.
The ESC exposed to 0, 19, 20, 50, and 100 μM of genistein, daidzein, and genistein + daidzein showed a dose-dependent proliferation decrease. After 48–96 hours of culture, this reduction was significant in the presence of 50 μM of phytoestrogens versus 10 μM untreated ESC. The ESC decidualized in the presence of phytoestrogens did not rearrange their cytoskeletons and showed a significant decrease in PRL secretion compared with untreated decidualized ESCs (dESCs). However, phytoestrogens did not alter proliferative status or the percentage of viable/apoptotic cells in dESC compared with untreated dESC. During decidualization, phytoestrogens induced the same nuclear translocation of ERβ and PR as the control dESC.
This study reveals that high doses of phytoestrogens could affect the in vitro decidualization process.
La exposición a fitoestrógenos altera células del estroma endometrial e interfiere con la señalización de la decidualización
Investigar si los fitoestrógenos (genisteína y daidzeína) alteran la decidualización de las células del estroma endometrial humano (CEE) in vitro.
Células estromales endometriales (CEE) primarias aisladas fueron expuestas a fitoestrógenos y decidualizadas in vitro.
Centro académico de fertilidad.
Veinte donantes de ovocitos fértiles que han acudido a la clínica IVI Valencia.
Tratamiento de las CEE con fitoestrógenos a 0, 10, 20, 50 y 100 μM.
La proliferación de CEE se analizó mediante el test MTS. La decidualización in vitro fue inducida en presencia de fitoestrógenos con acetato de medroxiprogesterona / adenosina cíclica monofosfato 3´:5´y evaluado por prolactina (PRL)-ELISA de prolactina (PRL) e inmunotinción actina F. El marcador proliferativo Ki67 fue analizado por inmunofluorescencia. La apoptosis de las CEE fue evaluada mediante la detección de anexina V / yoduro de propidio mediante citometría de flujo. La localización de los receptores de estrógeno (ERβ) y del receptor P de progesterona (PR) fue evaluado por inmunofluorescencia.
Las CEE expuestas a 0, 19, 20, 50 y 100 mM de genisteína, daidzeína y genisteína y daidzeína mostraron una disminución de la proliferación dosis dependiente. Después de 48-96 horas de cultivo, esta reducción fue significativa en presencia de 50 mM de fitoestrógenos versus 10 μM de CEE sin tratar. Las CEE decidualizadas en presencia de los fitoestrógenos no reorganizaron sus citoesqueletos y mostraron una disminución significativa en la secreción de PRL en comparación con las CEE decidualizados (dCEE) no tratadas. Sin embargo, los fitoestrógenos no alteraron el estado proliferativo o el porcentaje de células viables / apoptóticas en dCEE en comparación con dCEE no tratadas. Durante la decidualización, los fitoestrógenos indujeron la misma translocación nuclear de ERβ y PR que en las dCEE control.
Este estudio revela que altas dosis de fitoestrógenos podrían afectar el proceso de decidualización in vitro.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>31371049</pmid><doi>10.1016/j.fertnstert.2019.06.014</doi><orcidid>https://orcid.org/0000-0002-7620-8081</orcidid><oa>free_for_read</oa></addata></record> |
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source | Elsevier ScienceDirect Journals Complete - AutoHoldings; MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
subjects | Cell Proliferation - drug effects Cell Proliferation - physiology Cells, Cultured Decidua - cytology Decidua - drug effects Decidua - physiology decidualization Dose-Response Relationship, Drug endocrine disruptors endometrium Endometrium - cytology Endometrium - drug effects Endometrium - physiology Female Genistein - pharmacology Humans Isoflavones - pharmacology Phytoestrogens Phytoestrogens - pharmacology Signal Transduction - drug effects Signal Transduction - physiology steroid receptors Stromal Cells - drug effects Stromal Cells - physiology |
title | Phytoestrogen exposure alters endometrial stromal cells and interferes with decidualization signaling |
url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-16T09%3A58%3A39IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Phytoestrogen%20exposure%20alters%20endometrial%20stromal%20cells%20and%C2%A0interferes%20with%20decidualization%20signaling&rft.jtitle=Fertility%20and%20sterility&rft.au=Salsano,%20Stefania&rft.date=2019-11&rft.volume=112&rft.issue=5&rft.spage=947&rft.epage=958.e3&rft.pages=947-958.e3&rft.issn=0015-0282&rft.eissn=1556-5653&rft_id=info:doi/10.1016/j.fertnstert.2019.06.014&rft_dat=%3Cproquest_cross%3E2268308608%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2268308608&rft_id=info:pmid/31371049&rft_els_id=S0015028219305473&rfr_iscdi=true |