{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T22:32:50Z","timestamp":1790548370361,"version":"4.1.0"},"reference-count":48,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2024,10,2]],"date-time":"2024-10-02T00:00:00Z","timestamp":1727827200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Nature Science Foundation of China","award":["32170862"],"award-info":[{"award-number":["32170862"]}]},{"name":"National Nature Science Foundation of China","award":["2019SK1012"],"award-info":[{"award-number":["2019SK1012"]}]},{"name":"National Nature Science Foundation of China","award":["2022XKQ0205"],"award-info":[{"award-number":["2022XKQ0205"]}]},{"name":"National Nature Science Foundation of China","award":["2023JC101"],"award-info":[{"award-number":["2023JC101"]}]},{"name":"Major Scientific and Technological Projects for Collaborative Prevention and Control of Birth Defect in Hunan Province","award":["32170862"],"award-info":[{"award-number":["32170862"]}]},{"name":"Major Scientific and Technological Projects for Collaborative Prevention and Control of Birth Defect in Hunan Province","award":["2019SK1012"],"award-info":[{"award-number":["2019SK1012"]}]},{"name":"Major Scientific and Technological Projects for Collaborative Prevention and Control of Birth Defect in Hunan Province","award":["2022XKQ0205"],"award-info":[{"award-number":["2022XKQ0205"]}]},{"name":"Major Scientific and Technological Projects for Collaborative Prevention and Control of Birth Defect in Hunan Province","award":["2023JC101"],"award-info":[{"award-number":["2023JC101"]}]},{"name":"Developmental Biology and Breeding","award":["32170862"],"award-info":[{"award-number":["32170862"]}]},{"name":"Developmental Biology and Breeding","award":["2019SK1012"],"award-info":[{"award-number":["2019SK1012"]}]},{"name":"Developmental Biology and Breeding","award":["2022XKQ0205"],"award-info":[{"award-number":["2022XKQ0205"]}]},{"name":"Developmental Biology and Breeding","award":["2023JC101"],"award-info":[{"award-number":["2023JC101"]}]},{"name":"Research Team for Reproduction Health and Translational Medicine of Hunan Normal University","award":["32170862"],"award-info":[{"award-number":["32170862"]}]},{"name":"Research Team for Reproduction Health and Translational Medicine of Hunan Normal University","award":["2019SK1012"],"award-info":[{"award-number":["2019SK1012"]}]},{"name":"Research Team for Reproduction Health and Translational Medicine of Hunan Normal University","award":["2022XKQ0205"],"award-info":[{"award-number":["2022XKQ0205"]}]},{"name":"Research Team for Reproduction Health and Translational Medicine of Hunan Normal University","award":["2023JC101"],"award-info":[{"award-number":["2023JC101"]}]},{"name":"Shanghai Key Laboratory of Reproductive Medicine","award":["32170862"],"award-info":[{"award-number":["32170862"]}]},{"name":"Shanghai Key Laboratory of Reproductive Medicine","award":["2019SK1012"],"award-info":[{"award-number":["2019SK1012"]}]},{"name":"Shanghai Key Laboratory of Reproductive Medicine","award":["2022XKQ0205"],"award-info":[{"award-number":["2022XKQ0205"]}]},{"name":"Shanghai Key Laboratory of Reproductive Medicine","award":["2023JC101"],"award-info":[{"award-number":["2023JC101"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Organoids play pivotal roles in uncovering the molecular mechanisms underlying organogenesis, intercellular communication, and high-throughput drug screening. Testicular organoids are essential for exploring the genetic and epigenetic regulation of spermatogenesis in vivo and the treatment of male infertility. However, the formation of testicular organoids with full spermatogenesis has not yet been achieved. In this study, neonatal mouse testicular cells were isolated by two-step enzymatic digestion, and they were combined with Matrigel and transplanted subcutaneously into nude mice. Histological examination (H&amp;E) staining and immunohistochemistry revealed that cell grafts assembled to form seminiferous tubules that contained spermatogonial stem cells (SSCs) and Sertoli cells, as illustrated by the co-expression of PLZF (a hallmark for SSCs) and SOX9 (a marker for Sertoli cells) as well as the co-expression of UCHL1 (a hallmark for SSCs) and SOX9, after 8 weeks of transplantation. At 10 weeks of transplantation, SSCs could proliferate and differentiate into spermatocytes as evidenced by the expression of PCNA, Ki67, c-Kit, SYCP3, \u03b3-HA2X, and MLH1. Notably, testicular organoids were seen, and spermatids were observed within the lumen of testicular organoids after 16 weeks of transplantation, as shown by the presence of TNP1 and ACROSIN (hallmarks for spermatids). Collectively, these results implicate that we successfully established testicular organoids with spermatogenesis in vivo. This study thus provides an excellent platform for unveiling the mechanisms underlying mammalian spermatogenesis, and it might offer valuable male gametes for treating male infertility.<\/jats:p>","DOI":"10.3390\/cells13191642","type":"journal-article","created":{"date-parts":[[2024,10,2]],"date-time":"2024-10-02T11:29:24Z","timestamp":1727868564000},"page":"1642","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Establishment and Characterization of Testis Organoids with Proliferation and Differentiation of Spermatogonial Stem Cells into Spermatocytes and Spermatids"],"prefix":"10.3390","volume":"13","author":[{"given":"Dong","family":"Zhang","sequence":"first","affiliation":[{"name":"Key Laboratory of Model Animals and Stem Cell Biology in Hunan Province, Hunan Normal University School of Medicine, Changsha 410013, China"},{"name":"Research Center of Reproduction and Translational Medicine of Hunan Province, Changsha 410008, China"},{"name":"Manufacture-Based Learning & Research Demonstration Center for Human Reproductive Health New Technology of Hunan Normal University, Changsha 410013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wencong","family":"Jin","sequence":"additional","affiliation":[{"name":"Key Laboratory of Model Animals and Stem Cell Biology in Hunan Province, Hunan Normal University School of Medicine, Changsha 410013, China"},{"name":"Research Center of Reproduction and Translational Medicine of Hunan Province, Changsha 410008, China"},{"name":"Manufacture-Based Learning & Research Demonstration Center for Human Reproductive Health New Technology of Hunan Normal University, Changsha 410013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yinghong","family":"Cui","sequence":"additional","affiliation":[{"name":"Key Laboratory of Model Animals and Stem Cell Biology in Hunan Province, Hunan Normal University School of Medicine, Changsha 410013, China"},{"name":"Research Center of Reproduction and Translational Medicine of Hunan Province, Changsha 410008, China"},{"name":"Manufacture-Based Learning & Research Demonstration Center for Human Reproductive Health New Technology of Hunan Normal University, Changsha 410013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zuping","family":"He","sequence":"additional","affiliation":[{"name":"Key Laboratory of Model Animals and Stem Cell Biology in Hunan Province, Hunan Normal University School of Medicine, Changsha 410013, China"},{"name":"Research Center of Reproduction and Translational Medicine of Hunan Province, Changsha 410008, China"},{"name":"Manufacture-Based Learning & Research Demonstration Center for Human Reproductive Health New Technology of Hunan Normal University, Changsha 410013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,10,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1038\/ncb3312","article-title":"Organoids as an in vitro model of human development and disease","volume":"18","author":"Fatehullah","year":"2016","journal-title":"Nat. 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