{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T07:06:38Z","timestamp":1789455998642,"version":"build-2803163510"},"reference-count":38,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2023,9,12]],"date-time":"2023-09-12T00:00:00Z","timestamp":1694476800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100004484","name":"Tehran University of Medical Sciences and Health Services","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004484","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Artificial Organs"],"published-print":{"date-parts":[[2023,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Purpose<\/jats:title><jats:p>Development of organoids using human primary testicular cells has remained a challenge due to the complexity of the mammalian testicular cytoarchitecture and culture methods. In this study, we generated testicular organoids derived from human primary testicular cells. Then, we evaluated the effect of stem cell factor (SCF) on cell differentiation and apoptosis in the testicular organoid model.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The testicular cells were harvested from the three brain\u2010dead donors. Human spermatogonial stem cells (SSCs) were characterized using immunocytochemistry (ICC), RT\u2010PCR and flow cytometry. Testicular organoids were generated from primary testicular cells by hanging drop culture method and were cultured in three groups: control group, experimental group 1 (treated FSH and retinoic acid (RA)), and experimental group 2 (treated FSH, RA and SCF), for five weeks. We assessed the expression of SCP3 (Synaptonemal Complex Protein 3) as a meiotic gene, PRM2 (Protamine 2) as a post\u2010meiotic marker and apoptotic genes of Bax (BCL2\u2010Associated X Protein) and Bcl\u20102 (B\u2010cell lymphoma 2), respectively by using RT\u2010qPCR. In addition, we identified the expression of PRM2 by immunohistochemistry (IHC).<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Relative expression of SCP3, PRM2 and Bcl\u20102 were highest in group 2 after five weeks of culture. In contrast, BAX expression level was lower in experimental group 2 in comparison with other groups. IHC analyses indicated the highest expression of PRM2 as a postmeiotic marker in group 2 in comparison to 2D culture and control groups but not find significant differences between experimental group 1 and experimental group 2 groups. Morphological evaluations revealed that organoids are compact spherical structures and in the peripheral region composed of uncharacterized elongated fibroblast\u2010like cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Our findings revealed that the testicular organoid culture system promote the spermatogonial stem cell (SSC) differentiation, especially in presence of SCF. Developed organoids are capable of recapitulating many important properties of a stem cell niche.<\/jats:p><\/jats:sec>","DOI":"10.1111\/aor.14643","type":"journal-article","created":{"date-parts":[[2023,9,12]],"date-time":"2023-09-12T09:12:39Z","timestamp":1694509959000},"page":"1818-1830","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Differentiation of human primary testicular cells in the presence of <scp>SCF<\/scp> using the organoid culture system"],"prefix":"10.1111","volume":"47","author":[{"given":"Aghbibi","family":"Nikmahzar","sequence":"first","affiliation":[{"name":"Department of Anatomy, School of Medicine Tehran University of Medical Sciences  Tehran Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Morteza","family":"Koruji","sequence":"additional","affiliation":[{"name":"Stem Cell and Regenerative Medicine Center &amp; Department of Anatomy School of Medicine, Iran University of Medical Sciences  Tehran Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mehrdad","family":"Jahanshahi","sequence":"additional","affiliation":[{"name":"Neuroscience Research Center, Department of Anatomy, Faculty of Medicine Golestan University of Medical Sciences  Gorgan Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Farnaz","family":"Khadivi","sequence":"additional","affiliation":[{"name":"Department of Anatomy, School of Medicine Shahrekord University of Medical Sciences  Shahrekord Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maryam","family":"Shabani","sequence":"additional","affiliation":[{"name":"Department of Anatomy, School of Medicine Tehran University of Medical Sciences  Tehran Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sanaz","family":"Dehghani","sequence":"additional","affiliation":[{"name":"Organ Procurement Unit, Sina Hospital Tehran University of Medical Sciences  Tehran Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mehdi","family":"Forouzesh","sequence":"additional","affiliation":[{"name":"Legal Medicine Organization of Iran Legal Medicine Research Center, Legal Medicine Organization  Tehran Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ayob","family":"Jabari","sequence":"additional","affiliation":[{"name":"Department of Anatomy Zahedan Medical University of Science  Zahedan Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Narjes","family":"Feizollahi","sequence":"additional","affiliation":[{"name":"Department of Anatomy, School of Medicine Tehran University of Medical Sciences  Tehran Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maryam","family":"Salem","sequence":"additional","affiliation":[{"name":"Department of Anatomy, School of Medicine Tehran University of Medical Sciences  Tehran Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nasrin","family":"Ghanami\u00a0Gashti","sequence":"additional","affiliation":[{"name":"Biomaterials Cluster, Bernal Institute University of Limerick  Limerick Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasaman","family":"Abbasi","sequence":"additional","affiliation":[{"name":"Program in Neuroscience, Center to Advance Chronic Pain Research, Department of Neural and Pain Sciences, School of Dentistry University of Maryland  Baltimore Maryland USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4310-0381","authenticated-orcid":false,"given":"Mehdi","family":"Abbasi","sequence":"additional","affiliation":[{"name":"Department of Anatomy, School of Medicine Tehran University of Medical Sciences  Tehran Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2023,9,12]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpcell.00120.2020"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature12517"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature14415"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature12271"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2016.006"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncb3047"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1093\/humupd\/dmx036"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stemcr.2016.11.012"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aaf9414"},{"key":"e_1_2_9_11_1","first-page":"167","article-title":"Wistar rats immature testicular tissue vitrification and heterotopic grafting","volume":"22","author":"Benvenutti L","year":"2018","journal-title":"JBRA Assist Reprod"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.beem.2018.10.003"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.rbmo.2019.05.007"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41418-017-0015-1"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1093\/biolre\/ioz053"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1002\/wsbm.1590"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1007\/7651_2017_48"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biomaterials.2017.03.025"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.3390\/cells10071696"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod.116.143446"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1088\/1758-5090\/ab9907"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.113.1.161"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1002\/mrd.22430"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1530\/REP-08-0537"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stemcr.2014.08.004"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10815-012-9817-8"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1001\/jama.2009.1689"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mex.2020.100966"},{"key":"e_1_2_9_29_1","doi-asserted-by":"publisher","DOI":"10.1002\/jbm.1252"},{"key":"e_1_2_9_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biomaterials.2012.03.001"},{"issue":"2","key":"e_1_2_9_31_1","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0149492","article-title":"Direct measurements of oxygen gradients in spheroid culture system using electron parametric resonance oximetry","volume":"11","author":"Hinderberger D","year":"2016","journal-title":"PLoS One"},{"key":"e_1_2_9_32_1","doi-asserted-by":"publisher","DOI":"10.1093\/molehr\/gax070"},{"key":"e_1_2_9_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.modgep.2007.10.007"},{"key":"e_1_2_9_34_1","doi-asserted-by":"publisher","DOI":"10.4161\/cc.7.24.7262"},{"key":"e_1_2_9_35_1","doi-asserted-by":"publisher","DOI":"10.1080\/13880209.2019.1693603"},{"key":"e_1_2_9_36_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1439-0272.2010.01135.x"},{"key":"e_1_2_9_37_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.91.16.7553"},{"key":"e_1_2_9_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.fertnstert.2015.12.020"},{"key":"e_1_2_9_39_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1247125"}],"container-title":["Artificial Organs"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/aor.14643","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,8]],"date-time":"2024-01-08T10:24:33Z","timestamp":1704709473000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/aor.14643"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,9,12]]},"references-count":38,"journal-issue":{"issue":"12","published-print":{"date-parts":[[2023,12]]}},"alternative-id":["10.1111\/aor.14643"],"URL":"https:\/\/doi.org\/10.1111\/aor.14643","archive":["Portico"],"relation":{},"ISSN":["0160-564X","1525-1594"],"issn-type":[{"value":"0160-564X","type":"print"},{"value":"1525-1594","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,9,12]]},"assertion":[{"value":"2023-04-11","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-08-09","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-09-12","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}