Human testis in organotypic culture: application for basic or clinical research
Identifieur interne : 000E45 ( Istex/Corpus ); précédent : 000E44; suivant : 000E46Human testis in organotypic culture: application for basic or clinical research
Auteurs : V. Roulet ; H. Denis ; C. Staub ; A. Le Tortorec ; B. Delaleu ; A. P. Satie ; J. J. Patard ; B. Je Gou ; N. Dejucq-RainsfordSource :
- Human Reproduction [ 0268-1161 ] ; 2006-06.
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- KwdEn :
Abstract
BACKGROUND: Over recent decades, recurring efforts have been devoted to developing testicular cell or tissue cultures for basic and clinical research. However, there remains much confusion, particularly concerning the fate of human germ cells in culture. OBJECTIVE: To reassess the status of human testicular cell types as well as the ability of germ cells to divide and differentiate in organotypic culture. METHODS: Human testicular fragments were maintained for 2 weeks in culture. The viability and functionality of testicular cells were assessed using light and electronic microscopy, apoptotic cell labelling, 5-bromo-2′-deoxyuridine (BrdU) incorporation, immunohistochemistry and quantitative PCR against specific cell markers. RESULTS: A gradual loss of meiotic and post-meiotic germ cells occurred throughout the culture period, irrespective of the presence of gonadotrophins. However, all germ cell types remained traceable for up to 16 days, some still dividing and differentiating at a rate compatible with the in vivo situation. Good maintenance of the general architecture of the explants associated with clearly quantifiable levels of several somatic cell markers was observed. CONCLUSION: Although this culture model is clearly unsuitable for preparing germ cells for therapeutic purposes, it does represent a most valuable tool for testing the effects of biological and chemical agents on testicular tissue.
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DOI: 10.1093/humrep/del018
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Roulet</name><affiliation><mods:affiliation>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</mods:affiliation></affiliation><affiliation><mods:affiliation>Université de Rennes 1,</mods:affiliation></affiliation></author><author><name sortKey="Denis, H" sort="Denis, H" uniqKey="Denis H" first="H." last="Denis">H. Denis</name><affiliation><mods:affiliation>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</mods:affiliation></affiliation><affiliation><mods:affiliation>Université de Rennes 1,</mods:affiliation></affiliation></author><author><name sortKey="Staub, C" sort="Staub, C" uniqKey="Staub C" first="C." last="Staub">C. Staub</name><affiliation><mods:affiliation>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</mods:affiliation></affiliation><affiliation><mods:affiliation>Université de Rennes 1,</mods:affiliation></affiliation></author><author><name sortKey="Le Tortorec, A" sort="Le Tortorec, A" uniqKey="Le Tortorec A" first="A." last="Le Tortorec">A. Le Tortorec</name><affiliation><mods:affiliation>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</mods:affiliation></affiliation><affiliation><mods:affiliation>Université de Rennes 1,</mods:affiliation></affiliation></author><author><name sortKey="Delaleu, B" sort="Delaleu, B" uniqKey="Delaleu B" first="B." last="Delaleu">B. Delaleu</name><affiliation><mods:affiliation>Service de microscopie électronique, INRA PRC, Tours and</mods:affiliation></affiliation></author><author><name sortKey="Satie, A P" sort="Satie, A P" uniqKey="Satie A" first="A. P." last="Satie">A. P. Satie</name><affiliation><mods:affiliation>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</mods:affiliation></affiliation><affiliation><mods:affiliation>Université de Rennes 1,</mods:affiliation></affiliation></author><author><name sortKey="Patard, J J" sort="Patard, J J" uniqKey="Patard J" first="J. J." last="Patard">J. J. Patard</name><affiliation><mods:affiliation>Service d’Urologie, CHUR Pontchaillou, Rennes, France</mods:affiliation></affiliation></author><author><name sortKey="Je Gou, B" sort="Je Gou, B" uniqKey="Je Gou B" first="B." last="Je Gou">B. Je Gou</name><affiliation><mods:affiliation>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</mods:affiliation></affiliation><affiliation><mods:affiliation>6To whom correspondence should be addressed at: INSERM U625-GERHM, Campus de Beaulieu, 35042 Rennes Cedex, France.</mods:affiliation></affiliation><affiliation><mods:affiliation>E-mail: bernard.jegou@rennes.inserm.fr</mods:affiliation></affiliation><affiliation><mods:affiliation>Université de Rennes 1,</mods:affiliation></affiliation></author><author><name sortKey="Dejucq Rainsford, N" sort="Dejucq Rainsford, N" uniqKey="Dejucq Rainsford N" first="N." last="Dejucq-Rainsford">N. Dejucq-Rainsford</name><affiliation><mods:affiliation>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</mods:affiliation></affiliation><affiliation><mods:affiliation>7To whom correspondence should be addressed at: INSERM U625-GERHM, Campus de Beaulieu, 35042 Rennes Cedex, France.</mods:affiliation></affiliation><affiliation><mods:affiliation>E-mail: nathalie.dejucq-rainsford@rennes.inserm.fr</mods:affiliation></affiliation><affiliation><mods:affiliation>Université de Rennes 1,</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Human Reproduction</title><title level="j" type="abbrev">Hum. Reprod.</title><idno type="ISSN">0268-1161</idno><idno type="eISSN">1460-2350</idno><imprint><publisher>Oxford University Press</publisher><date type="published" when="2006-06">2006-06</date><biblScope unit="volume">21</biblScope><biblScope unit="issue">6</biblScope><biblScope unit="page" from="1564">1564</biblScope><biblScope unit="page" to="1575">1575</biblScope></imprint><idno type="ISSN">0268-1161</idno></series><idno type="istex">84604146EBCF09F64ECBACBCBB13D22C8B28D26E</idno><idno type="DOI">10.1093/humrep/del018</idno><idno type="local">018</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0268-1161</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>human</term><term>organotypic culture</term><term>testis</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">BACKGROUND: Over recent decades, recurring efforts have been devoted to developing testicular cell or tissue cultures for basic and clinical research. However, there remains much confusion, particularly concerning the fate of human germ cells in culture. OBJECTIVE: To reassess the status of human testicular cell types as well as the ability of germ cells to divide and differentiate in organotypic culture. METHODS: Human testicular fragments were maintained for 2 weeks in culture. The viability and functionality of testicular cells were assessed using light and electronic microscopy, apoptotic cell labelling, 5-bromo-2′-deoxyuridine (BrdU) incorporation, immunohistochemistry and quantitative PCR against specific cell markers. RESULTS: A gradual loss of meiotic and post-meiotic germ cells occurred throughout the culture period, irrespective of the presence of gonadotrophins. However, all germ cell types remained traceable for up to 16 days, some still dividing and differentiating at a rate compatible with the in vivo situation. Good maintenance of the general architecture of the explants associated with clearly quantifiable levels of several somatic cell markers was observed. CONCLUSION: Although this culture model is clearly unsuitable for preparing germ cells for therapeutic purposes, it does represent a most valuable tool for testing the effects of biological and chemical agents on testicular tissue.</div></front></TEI><istex><corpusName>oup</corpusName><author><json:item><name>V. Roulet</name><affiliations><json:string>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</json:string><json:string>Université de Rennes 1,</json:string></affiliations></json:item><json:item><name>H. Denis</name><affiliations><json:string>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</json:string><json:string>Université de Rennes 1,</json:string></affiliations></json:item><json:item><name>C. Staub</name><affiliations><json:string>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</json:string><json:string>Université de Rennes 1,</json:string></affiliations></json:item><json:item><name>A. Le Tortorec</name><affiliations><json:string>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</json:string><json:string>Université de Rennes 1,</json:string></affiliations></json:item><json:item><name>B. Delaleu</name><affiliations><json:string>Service de microscopie électronique, INRA PRC, Tours and</json:string></affiliations></json:item><json:item><name>A.P. Satie</name><affiliations><json:string>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</json:string><json:string>Université de Rennes 1,</json:string></affiliations></json:item><json:item><name>J.J. Patard</name><affiliations><json:string>Service d’Urologie, CHUR Pontchaillou, Rennes, France</json:string></affiliations></json:item><json:item><name>B. Jégou</name><affiliations><json:string>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</json:string><json:string>6To whom correspondence should be addressed at: INSERM U625-GERHM, Campus de Beaulieu, 35042 Rennes Cedex, France.</json:string><json:string>E-mail: bernard.jegou@rennes.inserm.fr</json:string><json:string>Université de Rennes 1,</json:string><json:string>E-mail: bernard.jegou@rennes.inserm.fr</json:string></affiliations></json:item><json:item><name>N. Dejucq-Rainsford</name><affiliations><json:string>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</json:string><json:string>7To whom correspondence should be addressed at: INSERM U625-GERHM, Campus de Beaulieu, 35042 Rennes Cedex, France.</json:string><json:string>E-mail: nathalie.dejucq-rainsford@rennes.inserm.fr</json:string><json:string>Université de Rennes 1,</json:string><json:string>E-mail: nathalie.dejucq-rainsford@rennes.inserm.fr</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>human</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>organotypic culture</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>testis</value></json:item></subject><language><json:string>eng</json:string></language><originalGenre><json:string>research-article</json:string></originalGenre><abstract>BACKGROUND: Over recent decades, recurring efforts have been devoted to developing testicular cell or tissue cultures for basic and clinical research. However, there remains much confusion, particularly concerning the fate of human germ cells in culture. OBJECTIVE: To reassess the status of human testicular cell types as well as the ability of germ cells to divide and differentiate in organotypic culture. METHODS: Human testicular fragments were maintained for 2 weeks in culture. The viability and functionality of testicular cells were assessed using light and electronic microscopy, apoptotic cell labelling, 5-bromo-2′-deoxyuridine (BrdU) incorporation, immunohistochemistry and quantitative PCR against specific cell markers. RESULTS: A gradual loss of meiotic and post-meiotic germ cells occurred throughout the culture period, irrespective of the presence of gonadotrophins. However, all germ cell types remained traceable for up to 16 days, some still dividing and differentiating at a rate compatible with the in vivo situation. Good maintenance of the general architecture of the explants associated with clearly quantifiable levels of several somatic cell markers was observed. CONCLUSION: Although this culture model is clearly unsuitable for preparing germ cells for therapeutic purposes, it does represent a most valuable tool for testing the effects of biological and chemical agents on testicular tissue.</abstract><qualityIndicators><score>7.424</score><pdfVersion>1.3</pdfVersion><pdfPageSize>612 x 791 pts</pdfPageSize><refBibsNative>true</refBibsNative><keywordCount>3</keywordCount><abstractCharCount>1424</abstractCharCount><pdfWordCount>8200</pdfWordCount><pdfCharCount>51290</pdfCharCount><pdfPageCount>12</pdfPageCount><abstractWordCount>202</abstractWordCount></qualityIndicators><title>Human testis in organotypic culture: application for basic or clinical research</title><genre><json:string>research-article</json:string></genre><host><volume>21</volume><publisherId><json:string>humrep</json:string></publisherId><pages><last>1575</last><first>1564</first></pages><issn><json:string>0268-1161</json:string></issn><issue>6</issue><genre><json:string>journal</json:string></genre><language><json:string>unknown</json:string></language><eissn><json:string>1460-2350</json:string></eissn><title>Human Reproduction</title></host><categories><wos><json:string>OBSTETRICS & GYNECOLOGY</json:string><json:string>REPRODUCTIVE BIOLOGY</json:string></wos></categories><publicationDate>2006</publicationDate><copyrightDate>2006</copyrightDate><doi><json:string>10.1093/humrep/del018</json:string></doi><id>84604146EBCF09F64ECBACBCBB13D22C8B28D26E</id><score>0.24937814</score><fulltext><json:item><original>true</original><mimetype>application/pdf</mimetype><extension>pdf</extension><uri>https://api.istex.fr/document/84604146EBCF09F64ECBACBCBB13D22C8B28D26E/fulltext/pdf</uri></json:item><json:item><original>false</original><mimetype>application/zip</mimetype><extension>zip</extension><uri>https://api.istex.fr/document/84604146EBCF09F64ECBACBCBB13D22C8B28D26E/fulltext/zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/document/84604146EBCF09F64ECBACBCBB13D22C8B28D26E/fulltext/tei"><teiHeader><fileDesc><titleStmt><title level="a" type="main" xml:lang="en">Human testis in organotypic culture: application for basic or clinical research</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher>Oxford University Press</publisher><availability><p>© The Author 2006. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org</p></availability><date>2006-02-23</date></publicationStmt><notesStmt><note>Submitted on April 15, 2005; resubmitted on June 30, 2005, November 29, 2005; accepted on January 10, 2006</note><note>6To whom correspondence should be addressed at: INSERM U625-GERHM, Campus de Beaulieu, 35042 Rennes Cedex, France. E-mail: bernard.jegou@rennes.inserm.fr</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a" type="main" xml:lang="en">Human testis in organotypic culture: application for basic or clinical research</title><author xml:id="author-1"><persName><forename type="first">V.</forename><surname>Roulet</surname></persName><affiliation>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</affiliation><affiliation>Université de Rennes 1,</affiliation></author><author xml:id="author-2"><persName><forename type="first">H.</forename><surname>Denis</surname></persName><affiliation>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</affiliation><affiliation>Université de Rennes 1,</affiliation></author><author xml:id="author-3"><persName><forename type="first">C.</forename><surname>Staub</surname></persName><affiliation>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</affiliation><affiliation>Université de Rennes 1,</affiliation></author><author xml:id="author-4"><persName><forename type="first">A.</forename><surname>Le Tortorec</surname></persName><affiliation>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</affiliation><affiliation>Université de Rennes 1,</affiliation></author><author xml:id="author-5"><persName><forename type="first">B.</forename><surname>Delaleu</surname></persName><affiliation>Service de microscopie électronique, INRA PRC, Tours and</affiliation></author><author xml:id="author-6"><persName><forename type="first">A.P.</forename><surname>Satie</surname></persName><affiliation>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</affiliation><affiliation>Université de Rennes 1,</affiliation></author><author xml:id="author-7"><persName><forename type="first">J.J.</forename><surname>Patard</surname></persName><affiliation>Service d’Urologie, CHUR Pontchaillou, Rennes, France</affiliation></author><author xml:id="author-8"><persName><forename type="first">B.</forename><surname>Jégou</surname></persName><email>bernard.jegou@rennes.inserm.fr</email><affiliation>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</affiliation><affiliation>6To whom correspondence should be addressed at: INSERM U625-GERHM, Campus de Beaulieu, 35042 Rennes Cedex, France.</affiliation><affiliation>Université de Rennes 1,</affiliation></author><author xml:id="author-9"><persName><forename type="first">N.</forename><surname>Dejucq-Rainsford</surname></persName><email>nathalie.dejucq-rainsford@rennes.inserm.fr</email><affiliation>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</affiliation><affiliation>7To whom correspondence should be addressed at: INSERM U625-GERHM, Campus de Beaulieu, 35042 Rennes Cedex, France.</affiliation><affiliation>Université de Rennes 1,</affiliation></author></analytic><monogr><title level="j">Human Reproduction</title><title level="j" type="abbrev">Hum. Reprod.</title><idno type="pISSN">0268-1161</idno><idno type="eISSN">1460-2350</idno><imprint><publisher>Oxford University Press</publisher><date type="published" when="2006-06"></date><biblScope unit="volume">21</biblScope><biblScope unit="issue">6</biblScope><biblScope unit="page" from="1564">1564</biblScope><biblScope unit="page" to="1575">1575</biblScope></imprint></monogr><idno type="istex">84604146EBCF09F64ECBACBCBB13D22C8B28D26E</idno><idno type="DOI">10.1093/humrep/del018</idno><idno type="local">018</idno></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>2006-02-23</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>BACKGROUND: Over recent decades, recurring efforts have been devoted to developing testicular cell or tissue cultures for basic and clinical research. However, there remains much confusion, particularly concerning the fate of human germ cells in culture. OBJECTIVE: To reassess the status of human testicular cell types as well as the ability of germ cells to divide and differentiate in organotypic culture. METHODS: Human testicular fragments were maintained for 2 weeks in culture. The viability and functionality of testicular cells were assessed using light and electronic microscopy, apoptotic cell labelling, 5-bromo-2′-deoxyuridine (BrdU) incorporation, immunohistochemistry and quantitative PCR against specific cell markers. RESULTS: A gradual loss of meiotic and post-meiotic germ cells occurred throughout the culture period, irrespective of the presence of gonadotrophins. However, all germ cell types remained traceable for up to 16 days, some still dividing and differentiating at a rate compatible with the in vivo situation. Good maintenance of the general architecture of the explants associated with clearly quantifiable levels of several somatic cell markers was observed. CONCLUSION: Although this culture model is clearly unsuitable for preparing germ cells for therapeutic purposes, it does represent a most valuable tool for testing the effects of biological and chemical agents on testicular tissue.</p></abstract><textClass xml:lang="en"><keywords scheme="keyword"><list><head>KWD</head><item><term>human</term></item><item><term>organotypic culture</term></item><item><term>testis</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="2006-02-23">Created</change><change when="2006-06">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><original>false</original><mimetype>text/plain</mimetype><extension>txt</extension><uri>https://api.istex.fr/document/84604146EBCF09F64ECBACBCBB13D22C8B28D26E/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="corpus oup" wicri:toSee="no header"><istex:xmlDeclaration>version="1.0" encoding="US-ASCII"</istex:xmlDeclaration><istex:docType PUBLIC="-//NLM//DTD Journal Publishing DTD v2.3 20070202//EN" URI="journalpublishing.dtd" name="istex:docType"></istex:docType><istex:document><article xml:lang="en" article-type="research-article"><front><journal-meta><journal-id journal-id-type="hwp">humrep</journal-id><journal-id journal-id-type="nlm-ta">Hum Reprod</journal-id><journal-id journal-id-type="publisher-id">humrep</journal-id><journal-title>Human Reproduction</journal-title><abbrev-journal-title abbrev-type="publisher">Hum. Reprod.</abbrev-journal-title><issn pub-type="ppub">0268-1161</issn><issn pub-type="epub">1460-2350</issn><publisher><publisher-name>Oxford University Press</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="other">018</article-id><article-id pub-id-type="doi">10.1093/humrep/del018</article-id><article-categories><subj-group subj-group-type="heading"><subject>Andrology</subject></subj-group></article-categories><title-group><article-title>Human testis in organotypic culture: application for basic or clinical research</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Roulet</surname><given-names>V.</given-names></name><xref rid="AFF1"><sup>1</sup></xref><xref rid="AFF2"><sup>2</sup></xref><xref rid="AFF3"><sup>3</sup></xref></contrib><contrib contrib-type="author"><name><surname>Denis</surname><given-names>H.</given-names></name><xref rid="AFF1"><sup>1</sup></xref><xref rid="AFF2"><sup>2</sup></xref><xref rid="AFF3"><sup>3</sup></xref></contrib><contrib contrib-type="author"><name><surname>Staub</surname><given-names>C.</given-names></name><xref rid="AFF1"><sup>1</sup></xref><xref rid="AFF2"><sup>2</sup></xref><xref rid="AFF3"><sup>3</sup></xref></contrib><contrib contrib-type="author"><name><surname>Le Tortorec</surname><given-names>A.</given-names></name><xref rid="AFF1"><sup>1</sup></xref><xref rid="AFF2"><sup>2</sup></xref><xref rid="AFF3"><sup>3</sup></xref></contrib><contrib contrib-type="author"><name><surname>Delaleu</surname><given-names>B.</given-names></name><xref rid="AFF4"><sup>4</sup></xref></contrib><contrib contrib-type="author"><name><surname>Satie</surname><given-names>A.P.</given-names></name><xref rid="AFF1"><sup>1</sup></xref><xref rid="AFF2"><sup>2</sup></xref><xref rid="AFF3"><sup>3</sup></xref></contrib><contrib contrib-type="author"><name><surname>Patard</surname><given-names>J.J.</given-names></name><xref rid="AFF5"><sup>5</sup></xref></contrib><contrib contrib-type="author"><name><surname>Jégou</surname><given-names>B.</given-names></name><xref rid="AFF1"><sup>1</sup></xref><xref rid="AFF2"><sup>2</sup></xref><xref rid="AFF3"><sup>3</sup></xref><xref rid="COR1"><sup>6</sup></xref></contrib><contrib contrib-type="author"><name><surname>Dejucq-Rainsford</surname><given-names>N.</given-names></name><xref rid="AFF1"><sup>1</sup></xref><xref rid="AFF2"><sup>2</sup></xref><xref rid="AFF3"><sup>3</sup></xref><xref rid="COR2"><sup>7</sup></xref></contrib><aff id="AFF1"><label>1</label>INSERM, U625, GERHM, Campus de Beaulieu, <target target-type="aff" id="AFF2"></target><label>2</label>Université de Rennes 1, <target target-type="aff" id="AFF3"></target><label>3</label>IFR 140, Rennes, <target target-type="aff" id="AFF4"></target><label>4</label>Service de microscopie électronique, INRA PRC, Tours and <target target-type="aff" id="AFF5"></target><label>5</label>Service d’Urologie, CHUR Pontchaillou, Rennes, France</aff></contrib-group><author-notes><corresp id="COR1"><label>6</label>To whom correspondence should be addressed at: INSERM U625-GERHM, Campus de Beaulieu, 35042 Rennes Cedex, France. E-mail: <ext-link xlink:href="bernard.jegou@rennes.inserm.fr" ext-link-type="email">bernard.jegou@rennes.inserm.fr</ext-link></corresp><corresp id="COR2"><label>7</label>To whom correspondence should be addressed at: INSERM U625-GERHM, Campus de Beaulieu, 35042 Rennes Cedex, France. E-mail: <ext-link xlink:href="nathalie.dejucq-rainsford@rennes.inserm.fr" ext-link-type="email">nathalie.dejucq-rainsford@rennes.inserm.fr</ext-link></corresp></author-notes><pub-date pub-type="epub"><day>23</day><month>2</month><year>2006</year></pub-date><pub-date pub-type="ppub"><month>June</month><year>2006</year></pub-date><volume>21</volume><issue>6</issue><fpage>1564</fpage><lpage>1575</lpage><permissions><copyright-statement>© The Author 2006. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org</copyright-statement><copyright-year>2006</copyright-year><license license-type="open-access"><p></p></license></permissions><abstract xml:lang="en"><p>BACKGROUND: Over recent decades, recurring efforts have been devoted to developing testicular cell or tissue cultures for basic and clinical research. However, there remains much confusion, particularly concerning the fate of human germ cells in culture. OBJECTIVE: To reassess the status of human testicular cell types as well as the ability of germ cells to divide and differentiate in organotypic culture. METHODS: Human testicular fragments were maintained for 2 weeks in culture. The viability and functionality of testicular cells were assessed using light and electronic microscopy, apoptotic cell labelling, 5-bromo-2′-deoxyuridine (BrdU) incorporation, immunohistochemistry and quantitative PCR against specific cell markers. RESULTS: A gradual loss of meiotic and post-meiotic germ cells occurred throughout the culture period, irrespective of the presence of gonadotrophins. However, all germ cell types remained traceable for up to 16 days, some still dividing and differentiating at a rate compatible with the <italic>in vivo</italic> situation. Good maintenance of the general architecture of the explants associated with clearly quantifiable levels of several somatic cell markers was observed. CONCLUSION: Although this culture model is clearly unsuitable for preparing germ cells for therapeutic purposes, it does represent a most valuable tool for testing the effects of biological and chemical agents on testicular tissue.</p></abstract><kwd-group kwd-group-type="KWD" xml:lang="en"><kwd>human</kwd><kwd>organotypic culture</kwd><kwd>testis</kwd></kwd-group><custom-meta-wrap><custom-meta><meta-name>hwp-legacy-fpage</meta-name><meta-value>1564</meta-value></custom-meta><custom-meta><meta-name>cover-date</meta-name><meta-value>June 2006</meta-value></custom-meta><custom-meta><meta-name>hwp-legacy-dochead</meta-name><meta-value>Article</meta-value></custom-meta></custom-meta-wrap></article-meta><notes><p content-type="arthw-misc"><italic>Submitted on April 15, 2005; resubmitted on June 30, 2005, November 29, 2005; accepted on January 10, 2006</italic></p></notes></front></article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Human testis in 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type="family">Jégou</namePart><affiliation>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</affiliation><affiliation>6To whom correspondence should be addressed at: INSERM U625-GERHM, Campus de Beaulieu, 35042 Rennes Cedex, France.</affiliation><affiliation>E-mail: bernard.jegou@rennes.inserm.fr</affiliation><affiliation>Université de Rennes 1,</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">N.</namePart><namePart type="family">Dejucq-Rainsford</namePart><affiliation>INSERM, U625, GERHM, Campus de Beaulieu, Université de Rennes 1, IFR 140, Rennes, Service de microscopie électronique, INRA PRC, Tours and Service d’Urologie, CHUR Pontchaillou, Rennes, France</affiliation><affiliation>7To whom correspondence should be addressed at: INSERM U625-GERHM, Campus de Beaulieu, 35042 Rennes Cedex, France.</affiliation><affiliation>E-mail: nathalie.dejucq-rainsford@rennes.inserm.fr</affiliation><affiliation>Université de Rennes 1,</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="research-article"></genre><originInfo><publisher>Oxford University Press</publisher><dateIssued encoding="w3cdtf">2006-06</dateIssued><dateCreated encoding="w3cdtf">2006-02-23</dateCreated><copyrightDate encoding="w3cdtf">2006</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType></physicalDescription><abstract lang="en">BACKGROUND: Over recent decades, recurring efforts have been devoted to developing testicular cell or tissue cultures for basic and clinical research. However, there remains much confusion, particularly concerning the fate of human germ cells in culture. OBJECTIVE: To reassess the status of human testicular cell types as well as the ability of germ cells to divide and differentiate in organotypic culture. METHODS: Human testicular fragments were maintained for 2 weeks in culture. The viability and functionality of testicular cells were assessed using light and electronic microscopy, apoptotic cell labelling, 5-bromo-2′-deoxyuridine (BrdU) incorporation, immunohistochemistry and quantitative PCR against specific cell markers. RESULTS: A gradual loss of meiotic and post-meiotic germ cells occurred throughout the culture period, irrespective of the presence of gonadotrophins. However, all germ cell types remained traceable for up to 16 days, some still dividing and differentiating at a rate compatible with the in vivo situation. Good maintenance of the general architecture of the explants associated with clearly quantifiable levels of several somatic cell markers was observed. CONCLUSION: Although this culture model is clearly unsuitable for preparing germ cells for therapeutic purposes, it does represent a most valuable tool for testing the effects of biological and chemical agents on testicular tissue.</abstract><note>Submitted on April 15, 2005; resubmitted on June 30, 2005, November 29, 2005; accepted on January 10, 2006</note><note type="author-notes">6To whom correspondence should be addressed at: INSERM U625-GERHM, Campus de Beaulieu, 35042 Rennes Cedex, France. E-mail: bernard.jegou@rennes.inserm.fr</note><subject lang="en"><genre>KWD</genre><topic>human</topic><topic>organotypic culture</topic><topic>testis</topic></subject><relatedItem type="host"><titleInfo><title>Human Reproduction</title></titleInfo><titleInfo type="abbreviated"><title>Hum. 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