Organotypic Culture of Testicular Tissue from Infant Boys with Cryptorchidism
Organotypic culture of human fetal testis has achieved fertilization-competent spermatids followed by blastocysts development. This study focuses on whether the organotypic culture of testicular tissue from infant boys with cryptorchidism could support the development of spermatogonia and somatic ce...
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| Veröffentlicht in: | International journal of molecular sciences 2022-07, Vol.23 (14), p.7975 |
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| creator | Wang, Danyang Hildorf, Simone Ntemou, Elissavet Mamsen, Linn Salto Dong, Lihua Pors, Susanne Elisabeth Fedder, Jens Clasen-Linde, Erik Cortes, Dina Thorup, Jørgen Andersen, Claus Yding |
| description | Organotypic culture of human fetal testis has achieved fertilization-competent spermatids followed by blastocysts development. This study focuses on whether the organotypic culture of testicular tissue from infant boys with cryptorchidism could support the development of spermatogonia and somatic cells. Frozen-thawed tissues were cultured in two different media, with or without retinoic acid (RA), for 60 days and evaluated by tissue morphology and immunostaining using germ and somatic cell markers. During the 60-day culture, spermatocytes stained by boule-like RNA-binding protein (BOLL) were induced in biopsies cultured with RA. Increased AR expression (p < 0.001) and decreased AMH expression (p < 0.001) in Sertoli cells indicated advancement of Sertoli cell maturity. An increased number of SOX9-positive Sertoli cells (p < 0.05) was observed, while the percentage of tubules with spermatogonia was reduced (p < 0.001). More tubules with alpha-smooth muscle actin (ACTA, peritubular myoid cells (PTMCs) marker) were observed in an RA-absent medium (p = 0.02). CYP17A1/STAR-positive Leydig cells demonstrated sustained steroidogenic function. Our culture conditions support the initiation of spermatocytes and enhanced maturation of Sertoli cells and PTMCs within infant testicular tissues. This study may be a basis for future studies focusing on maintaining and increasing the number of spermatogonia and identifying different factors and hormones, further advancing in vitro spermatogenesis. |
| doi_str_mv | 10.3390/ijms23147975 |
| format | Article |
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This study focuses on whether the organotypic culture of testicular tissue from infant boys with cryptorchidism could support the development of spermatogonia and somatic cells. Frozen-thawed tissues were cultured in two different media, with or without retinoic acid (RA), for 60 days and evaluated by tissue morphology and immunostaining using germ and somatic cell markers. During the 60-day culture, spermatocytes stained by boule-like RNA-binding protein (BOLL) were induced in biopsies cultured with RA. Increased AR expression (p < 0.001) and decreased AMH expression (p < 0.001) in Sertoli cells indicated advancement of Sertoli cell maturity. An increased number of SOX9-positive Sertoli cells (p < 0.05) was observed, while the percentage of tubules with spermatogonia was reduced (p < 0.001). More tubules with alpha-smooth muscle actin (ACTA, peritubular myoid cells (PTMCs) marker) were observed in an RA-absent medium (p = 0.02). CYP17A1/STAR-positive Leydig cells demonstrated sustained steroidogenic function. Our culture conditions support the initiation of spermatocytes and enhanced maturation of Sertoli cells and PTMCs within infant testicular tissues. This study may be a basis for future studies focusing on maintaining and increasing the number of spermatogonia and identifying different factors and hormones, further advancing in vitro spermatogenesis.</description><identifier>ISSN: 1422-0067</identifier><identifier>ISSN: 1661-6596</identifier><identifier>EISSN: 1422-0067</identifier><identifier>DOI: 10.3390/ijms23147975</identifier><identifier>PMID: 35887314</identifier><language>eng</language><publisher>Switzerland: MDPI AG</publisher><subject>Actin ; Antigens ; Biopsy ; Blastocysts ; Cell culture ; Cell growth ; Cryptorchidism ; Cryptorchidism - metabolism ; Cytology ; Fertility ; Fertilization ; Fetuses ; Gametocytes ; Growth factors ; Hormones ; Humans ; Infant ; Infants ; Leydig cells ; Male ; Markers ; Muscles ; Patients ; Retinoic acid ; RNA-binding protein ; Sertoli cells ; Sertoli Cells - metabolism ; Smooth muscle ; Somatic cells ; Sox9 protein ; Sperm ; Spermatids ; Spermatocytes ; Spermatogenesis ; Spermatogenesis - physiology ; Spermatogonia ; Spermatogonia - metabolism ; Surgery ; Testes ; Testis - metabolism ; Testosterone ; Tissue culture ; Tubules</subject><ispartof>International journal of molecular sciences, 2022-07, Vol.23 (14), p.7975</ispartof><rights>2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2022 by the authors. 2022</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c412t-c38a86a0d723d9cb2c57de69c5300af0b2ef8a17605b0a9e8e9f106d805a2ba33</citedby><cites>FETCH-LOGICAL-c412t-c38a86a0d723d9cb2c57de69c5300af0b2ef8a17605b0a9e8e9f106d805a2ba33</cites><orcidid>0000-0001-5383-5501 ; 0000-0002-7922-7034 ; 0000-0002-0130-1468</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9316019/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9316019/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,881,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35887314$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wang, Danyang</creatorcontrib><creatorcontrib>Hildorf, Simone</creatorcontrib><creatorcontrib>Ntemou, Elissavet</creatorcontrib><creatorcontrib>Mamsen, Linn Salto</creatorcontrib><creatorcontrib>Dong, Lihua</creatorcontrib><creatorcontrib>Pors, Susanne Elisabeth</creatorcontrib><creatorcontrib>Fedder, Jens</creatorcontrib><creatorcontrib>Clasen-Linde, Erik</creatorcontrib><creatorcontrib>Cortes, Dina</creatorcontrib><creatorcontrib>Thorup, Jørgen</creatorcontrib><creatorcontrib>Andersen, Claus Yding</creatorcontrib><title>Organotypic Culture of Testicular Tissue from Infant Boys with Cryptorchidism</title><title>International journal of molecular sciences</title><addtitle>Int J Mol Sci</addtitle><description>Organotypic culture of human fetal testis has achieved fertilization-competent spermatids followed by blastocysts development. This study focuses on whether the organotypic culture of testicular tissue from infant boys with cryptorchidism could support the development of spermatogonia and somatic cells. Frozen-thawed tissues were cultured in two different media, with or without retinoic acid (RA), for 60 days and evaluated by tissue morphology and immunostaining using germ and somatic cell markers. During the 60-day culture, spermatocytes stained by boule-like RNA-binding protein (BOLL) were induced in biopsies cultured with RA. Increased AR expression (p < 0.001) and decreased AMH expression (p < 0.001) in Sertoli cells indicated advancement of Sertoli cell maturity. An increased number of SOX9-positive Sertoli cells (p < 0.05) was observed, while the percentage of tubules with spermatogonia was reduced (p < 0.001). More tubules with alpha-smooth muscle actin (ACTA, peritubular myoid cells (PTMCs) marker) were observed in an RA-absent medium (p = 0.02). CYP17A1/STAR-positive Leydig cells demonstrated sustained steroidogenic function. Our culture conditions support the initiation of spermatocytes and enhanced maturation of Sertoli cells and PTMCs within infant testicular tissues. This study may be a basis for future studies focusing on maintaining and increasing the number of spermatogonia and identifying different factors and hormones, further advancing in vitro spermatogenesis.</description><subject>Actin</subject><subject>Antigens</subject><subject>Biopsy</subject><subject>Blastocysts</subject><subject>Cell culture</subject><subject>Cell growth</subject><subject>Cryptorchidism</subject><subject>Cryptorchidism - metabolism</subject><subject>Cytology</subject><subject>Fertility</subject><subject>Fertilization</subject><subject>Fetuses</subject><subject>Gametocytes</subject><subject>Growth factors</subject><subject>Hormones</subject><subject>Humans</subject><subject>Infant</subject><subject>Infants</subject><subject>Leydig cells</subject><subject>Male</subject><subject>Markers</subject><subject>Muscles</subject><subject>Patients</subject><subject>Retinoic acid</subject><subject>RNA-binding protein</subject><subject>Sertoli cells</subject><subject>Sertoli Cells - metabolism</subject><subject>Smooth muscle</subject><subject>Somatic cells</subject><subject>Sox9 protein</subject><subject>Sperm</subject><subject>Spermatids</subject><subject>Spermatocytes</subject><subject>Spermatogenesis</subject><subject>Spermatogenesis - physiology</subject><subject>Spermatogonia</subject><subject>Spermatogonia - metabolism</subject><subject>Surgery</subject><subject>Testes</subject><subject>Testis - metabolism</subject><subject>Testosterone</subject><subject>Tissue culture</subject><subject>Tubules</subject><issn>1422-0067</issn><issn>1661-6596</issn><issn>1422-0067</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>BENPR</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNpdkc1LAzEQxYMoWqs3zxLw4sHqJNlsNhdBix-FSi_1HLLZrE3Z3dRkV-l_75aqVE8zML_3mMdD6IzANWMSbtyyjpSRREjB99CAJJSOAFKxv7MfoeMYlwCUUS4P0RHjWSZ6zQC9zMKbbny7XjmDx13VdsFiX-K5ja0zXaUDnrsYO4vL4Gs8aUrdtPjeryP-dO0Cj8N61fpgFq5wsT5BB6Wuoj39nkP0-vgwHz-PprOnyfhuOjIJoe3IsExnqYZCUFZIk1PDRWFTaTgD0CXk1JaZJiIFnoOWNrOyJJAWGXBNc83YEN1ufVddXtvC2KYNulKr4God1sprp_5eGrdQb_5DSUZSILI3uPw2CP6967Oq2kVjq0o31ndR0VRyKhNJRI9e_EOXvgtNH29DJUAE52lPXW0pE3yMwZa_zxBQm57Ubk89fr4b4Bf-KYZ9AYN8j8A</recordid><startdate>20220719</startdate><enddate>20220719</enddate><creator>Wang, Danyang</creator><creator>Hildorf, Simone</creator><creator>Ntemou, Elissavet</creator><creator>Mamsen, Linn Salto</creator><creator>Dong, Lihua</creator><creator>Pors, Susanne Elisabeth</creator><creator>Fedder, Jens</creator><creator>Clasen-Linde, Erik</creator><creator>Cortes, Dina</creator><creator>Thorup, Jørgen</creator><creator>Andersen, Claus Yding</creator><general>MDPI AG</general><general>MDPI</general><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>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8G5</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>GUQSH</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>M2O</scope><scope>MBDVC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0001-5383-5501</orcidid><orcidid>https://orcid.org/0000-0002-7922-7034</orcidid><orcidid>https://orcid.org/0000-0002-0130-1468</orcidid></search><sort><creationdate>20220719</creationdate><title>Organotypic Culture of Testicular Tissue from Infant Boys with Cryptorchidism</title><author>Wang, Danyang ; Hildorf, Simone ; Ntemou, Elissavet ; Mamsen, Linn Salto ; Dong, Lihua ; Pors, Susanne Elisabeth ; Fedder, Jens ; Clasen-Linde, Erik ; Cortes, Dina ; Thorup, Jørgen ; Andersen, Claus Yding</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c412t-c38a86a0d723d9cb2c57de69c5300af0b2ef8a17605b0a9e8e9f106d805a2ba33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Actin</topic><topic>Antigens</topic><topic>Biopsy</topic><topic>Blastocysts</topic><topic>Cell culture</topic><topic>Cell growth</topic><topic>Cryptorchidism</topic><topic>Cryptorchidism - metabolism</topic><topic>Cytology</topic><topic>Fertility</topic><topic>Fertilization</topic><topic>Fetuses</topic><topic>Gametocytes</topic><topic>Growth factors</topic><topic>Hormones</topic><topic>Humans</topic><topic>Infant</topic><topic>Infants</topic><topic>Leydig cells</topic><topic>Male</topic><topic>Markers</topic><topic>Muscles</topic><topic>Patients</topic><topic>Retinoic acid</topic><topic>RNA-binding protein</topic><topic>Sertoli cells</topic><topic>Sertoli Cells - metabolism</topic><topic>Smooth muscle</topic><topic>Somatic cells</topic><topic>Sox9 protein</topic><topic>Sperm</topic><topic>Spermatids</topic><topic>Spermatocytes</topic><topic>Spermatogenesis</topic><topic>Spermatogenesis - physiology</topic><topic>Spermatogonia</topic><topic>Spermatogonia - metabolism</topic><topic>Surgery</topic><topic>Testes</topic><topic>Testis - metabolism</topic><topic>Testosterone</topic><topic>Tissue culture</topic><topic>Tubules</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wang, Danyang</creatorcontrib><creatorcontrib>Hildorf, Simone</creatorcontrib><creatorcontrib>Ntemou, Elissavet</creatorcontrib><creatorcontrib>Mamsen, Linn Salto</creatorcontrib><creatorcontrib>Dong, Lihua</creatorcontrib><creatorcontrib>Pors, Susanne Elisabeth</creatorcontrib><creatorcontrib>Fedder, Jens</creatorcontrib><creatorcontrib>Clasen-Linde, Erik</creatorcontrib><creatorcontrib>Cortes, Dina</creatorcontrib><creatorcontrib>Thorup, Jørgen</creatorcontrib><creatorcontrib>Andersen, Claus Yding</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Research Library (Alumni Edition)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>Research Library Prep</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Research Library</collection><collection>Research Library (Corporate)</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>International journal of molecular sciences</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wang, Danyang</au><au>Hildorf, Simone</au><au>Ntemou, Elissavet</au><au>Mamsen, Linn Salto</au><au>Dong, Lihua</au><au>Pors, Susanne Elisabeth</au><au>Fedder, Jens</au><au>Clasen-Linde, Erik</au><au>Cortes, Dina</au><au>Thorup, Jørgen</au><au>Andersen, Claus Yding</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Organotypic Culture of Testicular Tissue from Infant Boys with Cryptorchidism</atitle><jtitle>International journal of molecular sciences</jtitle><addtitle>Int J Mol Sci</addtitle><date>2022-07-19</date><risdate>2022</risdate><volume>23</volume><issue>14</issue><spage>7975</spage><pages>7975-</pages><issn>1422-0067</issn><issn>1661-6596</issn><eissn>1422-0067</eissn><abstract>Organotypic culture of human fetal testis has achieved fertilization-competent spermatids followed by blastocysts development. This study focuses on whether the organotypic culture of testicular tissue from infant boys with cryptorchidism could support the development of spermatogonia and somatic cells. Frozen-thawed tissues were cultured in two different media, with or without retinoic acid (RA), for 60 days and evaluated by tissue morphology and immunostaining using germ and somatic cell markers. During the 60-day culture, spermatocytes stained by boule-like RNA-binding protein (BOLL) were induced in biopsies cultured with RA. Increased AR expression (p < 0.001) and decreased AMH expression (p < 0.001) in Sertoli cells indicated advancement of Sertoli cell maturity. An increased number of SOX9-positive Sertoli cells (p < 0.05) was observed, while the percentage of tubules with spermatogonia was reduced (p < 0.001). More tubules with alpha-smooth muscle actin (ACTA, peritubular myoid cells (PTMCs) marker) were observed in an RA-absent medium (p = 0.02). CYP17A1/STAR-positive Leydig cells demonstrated sustained steroidogenic function. Our culture conditions support the initiation of spermatocytes and enhanced maturation of Sertoli cells and PTMCs within infant testicular tissues. This study may be a basis for future studies focusing on maintaining and increasing the number of spermatogonia and identifying different factors and hormones, further advancing in vitro spermatogenesis.</abstract><cop>Switzerland</cop><pub>MDPI AG</pub><pmid>35887314</pmid><doi>10.3390/ijms23147975</doi><orcidid>https://orcid.org/0000-0001-5383-5501</orcidid><orcidid>https://orcid.org/0000-0002-7922-7034</orcidid><orcidid>https://orcid.org/0000-0002-0130-1468</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Actin Antigens Biopsy Blastocysts Cell culture Cell growth Cryptorchidism Cryptorchidism - metabolism Cytology Fertility Fertilization Fetuses Gametocytes Growth factors Hormones Humans Infant Infants Leydig cells Male Markers Muscles Patients Retinoic acid RNA-binding protein Sertoli cells Sertoli Cells - metabolism Smooth muscle Somatic cells Sox9 protein Sperm Spermatids Spermatocytes Spermatogenesis Spermatogenesis - physiology Spermatogonia Spermatogonia - metabolism Surgery Testes Testis - metabolism Testosterone Tissue culture Tubules |
| title | Organotypic Culture of Testicular Tissue from Infant Boys with Cryptorchidism |
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