{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T04:22:31Z","timestamp":1790050951333,"version":"4.0.1"},"reference-count":82,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2024,8,5]],"date-time":"2024-08-05T00:00:00Z","timestamp":1722816000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Project of the Natural Science Foundation of Guangdong Province, China","award":["2020A1515110362"],"award-info":[{"award-number":["2020A1515110362"]}]},{"name":"Project of the Natural Science Foundation of Guangdong Province, China","award":["2021E05028"],"award-info":[{"award-number":["2021E05028"]}]},{"name":"Project of the Natural Science Foundation of Guangdong Province, China","award":["K23438"],"award-info":[{"award-number":["K23438"]}]},{"name":"Marine Biology Foundation of Zhanjiang Bureau of Science and Technology","award":["2020A1515110362"],"award-info":[{"award-number":["2020A1515110362"]}]},{"name":"Marine Biology Foundation of Zhanjiang Bureau of Science and Technology","award":["2021E05028"],"award-info":[{"award-number":["2021E05028"]}]},{"name":"Marine Biology Foundation of Zhanjiang Bureau of Science and Technology","award":["K23438"],"award-info":[{"award-number":["K23438"]}]},{"name":"International Exchange and Cooperation Project of Science and Technology Department of Guangdong Province","award":["2020A1515110362"],"award-info":[{"award-number":["2020A1515110362"]}]},{"name":"International Exchange and Cooperation Project of Science and Technology Department of Guangdong Province","award":["2021E05028"],"award-info":[{"award-number":["2021E05028"]}]},{"name":"International Exchange and Cooperation Project of Science and Technology Department of Guangdong Province","award":["K23438"],"award-info":[{"award-number":["K23438"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Sperm, a crucial gamete for reproduction in sexual reproduction, is generated through the proliferation, differentiation, and morphological transformations of spermatogonial stem cells within the specialized microenvironment of the testes. Replicating this environment artificially presents challenges. However, interdisciplinary advancements in physics, materials science, and cell engineering have facilitated the utilization of innovative materials, technologies, and structures for inducing in vitro sperm production. This article offers a comprehensive overview of research progress on inducing in vitro sperm production by categorizing techniques into two major systems based on matrix-based and non-matrix-based approaches, respectively. Detailed discussions are provided for both types of technology systems through comparisons of their similarities and differences, as well as research advancements. The aim is to provide researchers in this field with a comprehensive panoramic view while presenting our own perspectives and prospects.<\/jats:p>","DOI":"10.3390\/ijms25158524","type":"journal-article","created":{"date-parts":[[2024,8,5]],"date-time":"2024-08-05T18:21:40Z","timestamp":1722882100000},"page":"8524","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Recent Progress of Induced Spermatogenesis In Vitro"],"prefix":"10.3390","volume":"25","author":[{"given":"Siqi","family":"Liu","sequence":"first","affiliation":[{"name":"College of Coastal Agriculture Science, Guangdong Ocean University, Zhanjiang 524088, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0771-1791","authenticated-orcid":false,"given":"Jiang","family":"Wu","sequence":"additional","affiliation":[{"name":"College of Coastal Agriculture Science, Guangdong Ocean University, Zhanjiang 524088, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7190-423X","authenticated-orcid":false,"given":"Xin","family":"Zhao","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Mechanical Behavior of Materials, Xi\u2019an Jiaotong University, Xi\u2019an 710049, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meng","family":"Yu","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Mechanical Behavior of Materials, Xi\u2019an Jiaotong University, Xi\u2019an 710049, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masayasu","family":"Taniguchi","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, Yamaguchi University, Yamaguchi 753-0841, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huimingda","family":"Bao","sequence":"additional","affiliation":[{"name":"College of Coastal Agriculture Science, Guangdong Ocean University, Zhanjiang 524088, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai","family":"Kang","sequence":"additional","affiliation":[{"name":"College of Coastal Agriculture Science, Guangdong Ocean University, Zhanjiang 524088, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,8,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"849","DOI":"10.1111\/andr.13386","article-title":"What are Sertoli cells? Historical, methodological, and functional aspects","volume":"11","author":"Thumfart","year":"2023","journal-title":"Andrology"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Liu, Y., Teng, Z., Wang, Z., Zhu, P., Wang, Z., Liu, F., and Liu, X. (2023). Human umbilical cord mesenchymal stem cells (hUC-MSCs) alleviate paclitaxel-induced spermatogenesis defects and maintain male fertility. Biol. Res., 56.","DOI":"10.1186\/s40659-023-00459-w"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1038\/nrurol.2017.69","article-title":"Oxidative stress and male infertility","volume":"14","author":"Bisht","year":"2017","journal-title":"Nat. Rev. Urol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"810","DOI":"10.1016\/j.fertnstert.2018.08.015","article-title":"Male infertility as a window to health","volume":"110","author":"Choy","year":"2018","journal-title":"Fertil. Steril."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1016\/j.ydbio.2017.10.002","article-title":"Retinoic acid deficiency leads to an increase in spermatogonial stem number in the neonatal mouse testis, but excess retinoic acid results in no change","volume":"432","author":"Agrimson","year":"2017","journal-title":"Dev. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Wu, J., Kang, K., Liu, S., Ma, Y., Yu, M., and Zhao, X. (2023). Recent Progress of In Vitro 3D Culture of Male Germ Stem Cells. J. Funct. Biomater., 14.","DOI":"10.3390\/jfb14110543"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"564","DOI":"10.1210\/er.2014-1101","article-title":"The Mammalian Blood-Testis Barrier: Its Biology and Regulation","volume":"36","author":"Mruk","year":"2015","journal-title":"Endocr. Rev."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"873","DOI":"10.1095\/biolreprod65.3.873","article-title":"In vitro production of haploid germ cells from fresh or frozen-thawed testicular cells of neonatal bulls","volume":"65","author":"Lee","year":"2001","journal-title":"Biol. Reprod."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1016\/j.fertnstert.2006.09.015","article-title":"In vitro differentiation of germ cells from nonobstructive azoospermic patients using three-dimensional culture in a collagen gel matrix","volume":"87","author":"Lee","year":"2007","journal-title":"Fertil. Steril."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1146\/annurev.cellbio.24.110707.175355","article-title":"Regulation of spermatogonial stem cell self-renewal in mammals","volume":"24","author":"Oatley","year":"2008","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"s27","DOI":"10.1017\/S1751731118000435","article-title":"Review: Spermatogenesis in the bull","volume":"12","author":"Staub","year":"2018","journal-title":"Animal"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"601","DOI":"10.1093\/molehr\/gaw047","article-title":"In vitro differentiation of rat spermatogonia into round spermatids in tissue culture","volume":"22","author":"Reda","year":"2016","journal-title":"Mol. Hum. Reprod."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1199","DOI":"10.1016\/S0093-691X(97)00353-1","article-title":"Factors affecting spermatogenesis in the stallion","volume":"48","author":"Johnson","year":"1997","journal-title":"Theriogenology"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Thonnes, M., Vogt, M., Steinborn, K., Hausken, K.N., Levavi-Sivan, B., Froschauer, A., and Pfennig, F. (2020). An ex vivo Approach to Study Hormonal Control of Spermatogenesis in the Teleost Oreochromis niloticus. Front. Endocrinol., 11.","DOI":"10.3389\/fendo.2020.00443"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1016\/j.repbio.2018.09.006","article-title":"Efficiency of colony formation and differentiation of human spermatogenic cells in two different culture systems","volume":"18","author":"Gholami","year":"2018","journal-title":"Reprod. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"343","DOI":"10.24272\/j.issn.2095-8137.2019.051","article-title":"Progress in in vitro culture and gene editing of porcine spermatogonial stem cells","volume":"40","author":"Sun","year":"2019","journal-title":"Zool. Res."},{"key":"ref_17","first-page":"17","article-title":"In vitro Culture of Human Testicular Stem Cells on Feeder-Free Condition","volume":"14","author":"Piravar","year":"2013","journal-title":"J. Reprod. Infertil."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"5","DOI":"10.4103\/aja.aja_41_21","article-title":"Recent advances in isolation, identification, and culture of mammalian spermatogonial stem cells","volume":"24","author":"Xi","year":"2022","journal-title":"Asian J. Androl."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1093\/molehr\/gaz043","article-title":"In-vitro differentiation of early pig spermatogenic cells to haploid germ cells","volume":"25","author":"Yu","year":"2019","journal-title":"Mol. Hum. Reprod."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/j.devcel.2007.01.002","article-title":"Functional identification of the actual and potential stem cell compartments in mouse spermatogenesis","volume":"12","author":"Nakagawa","year":"2007","journal-title":"Dev. Cell"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"658","DOI":"10.1016\/j.stem.2014.01.019","article-title":"Mouse spermatogenic stem cells continually interconvert between equipotent singly isolated and syncytial states","volume":"14","author":"Hara","year":"2014","journal-title":"Cell Stem Cell"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"344","DOI":"10.1017\/S0967199421000733","article-title":"Evaluation of co-cultured spermatogonial stem cells encapsulated in alginate hydrogel with Sertoli cells and their transplantation into azoospermic mice","volume":"30","author":"Veisi","year":"2022","journal-title":"Zygote"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1121","DOI":"10.1089\/scd.2017.0018","article-title":"Long-Term Propagation of Porcine Undifferentiated Spermatogonia","volume":"26","author":"Zhang","year":"2017","journal-title":"Stem Cells Dev."},{"key":"ref_24","first-page":"79","article-title":"Role of Somatic Testicular Cells during Mouse Spermatogenesis in Three-Dimensional Collagen Gel Culture System","volume":"16","author":"Khajavi","year":"2014","journal-title":"Cell J."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"985","DOI":"10.1095\/biolreprod.104.036400","article-title":"Long-term culture of mouse male germline stem cells under serum-or feeder-free conditions","volume":"72","author":"Miki","year":"2005","journal-title":"Biol. Reprod."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1262\/jrd.2012-130","article-title":"Characterization and in vitro culture of putative spermatogonial stem cells derived from feline testicular tissue","volume":"59","author":"Tiptanavattana","year":"2013","journal-title":"J. Reprod. Dev."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2614","DOI":"10.2741\/1995","article-title":"In vitro propagation of spermatogonial stem cells from KM mice","volume":"11","author":"Shi","year":"2006","journal-title":"Front. Biosci."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Oatley, M.J., Kaucher, A.V., Yang, Q.E., Waqas, M.S., and Oatley, J.M. (2016). Conditions for Long-Term Culture of Cattle Undifferentiated Spermatogonia. Biol. Reprod., 95.","DOI":"10.1095\/biolreprod.116.139832"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1186\/s13287-021-02295-9","article-title":"Mesenchymal stromal\/stem cells and their exosomes for restoration of spermatogenesis in non-obstructive azoospermia: A systemic review","volume":"12","author":"Zhankina","year":"2021","journal-title":"Stem Cell Res. Ther."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1186\/s13287-021-02275-z","article-title":"Human placental mesenchymal stem cells ameliorate chemotherapy-induced damage in the testis by reducing apoptosis\/oxidative stress and promoting autophagy","volume":"12","author":"Lu","year":"2021","journal-title":"Stem Cell Res. Ther."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Onen, S., Kose, S., Yersal, N., and Korkusuz, P. (2022). Mesenchymal stem cells promote spermatogonial stem\/progenitor cell pool and spermatogenesis in neonatal mice in vitro. Sci. Rep., 12.","DOI":"10.1038\/s41598-022-15358-5"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1186\/s13287-023-03356-x","article-title":"A pumpless monolayer microfluidic device based on mesenchymal stem cell-conditioned medium promotes neonatal mouse in vitro spermatogenesis","volume":"14","author":"Onen","year":"2023","journal-title":"Stem Cell Res. Ther."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1093\/humrep\/deu330","article-title":"Derivation and characterization of a cytocompatible scaffold from human testis","volume":"30","author":"Baert","year":"2015","journal-title":"Hum. Reprod."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2197","DOI":"10.1007\/s11033-021-07041-1","article-title":"Effect of NMDA receptor agonist and antagonist on spermatogonial stem cells proliferation in 2- and 3- dimensional culture systems","volume":"49","author":"Noghani","year":"2022","journal-title":"Mol. Biol. Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1016\/j.theriogenology.2022.08.004","article-title":"Differentiation of neonate mouse spermatogonia on two-dimensional and three-dimensional culture systems supplemented with d-Serine and Dizocilpine (MK-801)","volume":"191","author":"Noghani","year":"2022","journal-title":"Theriogenology"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"151623","DOI":"10.1016\/j.acthis.2020.151623","article-title":"Isolation, identification and differentiation of human spermatogonial cells on three-dimensional decellularized sheep testis","volume":"122","author":"Ashouri","year":"2020","journal-title":"Acta Histochem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1016\/j.ijbiomac.2022.07.127","article-title":"In vitro production of mouse morphological sperm in artificial testis bioengineered by 3D printing of extracellular matrix","volume":"217","author":"Bashiri","year":"2022","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e1565","DOI":"10.31661\/gmj.v8i0.1565","article-title":"In Vitro Spermatogenesis by Three-dimensional Culture of Spermatogonial Stem Cells on Decellularized Testicular Matrix","volume":"8","author":"Ashouri","year":"2019","journal-title":"Galen. Med. J."},{"key":"ref_39","first-page":"410","article-title":"Three-Dimensional Culture of Mouse Spermatogonial Stem Cells Using a Decellularised Testicular Scaffold","volume":"21","author":"Majidi","year":"2020","journal-title":"Cell J."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"121057","DOI":"10.1016\/j.lfs.2022.121057","article-title":"Improving the process of spermatogenesis in azoospermic mice using spermatogonial stem cells co-cultured with epididymosomes in three-dimensional culture system","volume":"310","author":"Rahbar","year":"2022","journal-title":"Life Sci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1585","DOI":"10.1080\/21691401.2019.1594854","article-title":"Transplantation of mouse iPSCs into testis of azoospermic mouse model: In vivo and in vitro study","volume":"47","author":"Rahmani","year":"2019","journal-title":"Artif. Cells Nanomed. Biotechnol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"151627","DOI":"10.1016\/j.acthis.2020.151627","article-title":"Application of platelet-rich plasma (PRP) improves self-renewal of human spermatogonial stem cells in two-dimensional and three-dimensional culture systems","volume":"122","author":"Khadivi","year":"2020","journal-title":"Acta Histochem."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1316","DOI":"10.1002\/cbin.10844","article-title":"Porcine spermatogonial stem cells self-renew effectively in a three dimensional culture microenvironment","volume":"41","author":"Park","year":"2017","journal-title":"Cell Biol. Int."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"AbuMadighem, A., Solomon, R., Stepanovsky, A., Kapelushnik, J., Shi, Q., Meese, E., Lunenfeld, E., and Huleihel, M. (2018). Development of Spermatogenesis In Vitro in Three-Dimensional Culture from Spermatogonial Cells of Busulfan-Treated Immature Mice. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19123804"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"972","DOI":"10.4103\/1008-682X.154994","article-title":"Application of three-dimensional culture systems to study mammalian spermatogenesis, with an emphasis on the rhesus monkey (Macaca mulatta)","volume":"17","author":"Huleihel","year":"2015","journal-title":"Asian J. Androl."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Abofoul-Azab, M., Lunenfeld, E., Levitas, E., Zeadna, A., Younis, J.S., Bar-Ami, S., and Huleihel, M. (2019). Identification of Premeiotic, Meiotic, and Postmeiotic Cells in Testicular Biopsies Without Sperm from Sertoli Cell-Only Syndrome Patients. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20030470"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1017\/S0967199421000551","article-title":"Use of alginate hydrogel to improve long-term 3D culture of spermatogonial stem cells: Stemness gene expression and structural features","volume":"30","author":"Hemadi","year":"2022","journal-title":"Zygote"},{"key":"ref_48","first-page":"4563","article-title":"The effects of poly L-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture","volume":"8","author":"Eslahi","year":"2013","journal-title":"Int. J. Nanomed."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1080\/19396368.2020.1725927","article-title":"Differentiation of neonate mouse spermatogonial stem cells on three-dimensional agar\/polyvinyl alcohol nanofiber scaffold","volume":"66","author":"Ziloochi","year":"2020","journal-title":"Syst. Biol. Reprod. Med."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"e14380","DOI":"10.1111\/and.14380","article-title":"Proliferation of human spermatogonial stem cells on optimized PCL\/Gelatin nanofibrous scaffolds","volume":"54","author":"Bashiri","year":"2022","journal-title":"Andrologia"},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Jabari, A., Gholami, K., Khadivi, F., Koruji, M., Amidi, F., Gilani, M., Mahabadi, V.P., Nikmahzar, A., Salem, M., and Movassagh, S.A. (2023). In vitro complete differentiation of human spermatogonial stem cells to morphologic spermatozoa using a hybrid hydrogel of agarose and laminin. Int. J. Biol. Macromol., 235.","DOI":"10.1016\/j.ijbiomac.2023.123801"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"151572","DOI":"10.1016\/j.acthis.2020.151572","article-title":"Three-dimensional co-culture of human spermatogonial stem cells with Sertoli cells in soft agar culture system supplemented by growth factors and Laminin","volume":"122","author":"Jabari","year":"2020","journal-title":"Acta Histochem."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1111\/rda.14043","article-title":"Isolation and in vitro expansion of porcine spermatogonial stem cells","volume":"57","author":"Zhao","year":"2022","journal-title":"Reprod. Domest. Anim."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1365","DOI":"10.1002\/jcp.24880","article-title":"An alternative long-term culture system for highly-pure mouse spermatogonial stem cells","volume":"230","author":"He","year":"2015","journal-title":"J. Cell Physiol."},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Xu, X., Feng, Q., Ma, X., Deng, Y., Zhang, K., Ooi, H.S., Yang, B., Zhang, Z.Y., Feng, B., and Bian, L. (2022). Dynamic gelatin-based hydrogels promote the proliferation and self-renewal of embryonic stem cells in long-term 3D culture. Biomaterials, 289.","DOI":"10.1016\/j.biomaterials.2022.121802"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.jconrel.2016.05.037","article-title":"Transplantation of testicular tissue in alginate hydrogel loaded with VEGF nanoparticles improves spermatogonial recovery","volume":"234","author":"Poels","year":"2016","journal-title":"J. Control Release"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1007\/s11626-020-00437-6","article-title":"The air-liquid interface culture of the mechanically isolated seminiferous tubules embedded in agarose or alginate improves in vitro spermatogenesis at the expense of attenuating their integrity","volume":"56","author":"Gholami","year":"2020","journal-title":"In Vitro Cell Dev. Biol. Anim."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1186\/s13287-018-0997-8","article-title":"Organ culture of seminiferous tubules using a modified soft agar culture system","volume":"9","author":"Gholami","year":"2018","journal-title":"Stem Cell Res. Ther."},{"key":"ref_59","doi-asserted-by":"crossref","unstructured":"Kojima, K., Sato, T., Naruse, Y., and Ogawa, T. (2016). Spermatogenesis in Explanted Fetal Mouse Testis Tissues. Biol. Reprod., 95.","DOI":"10.1095\/biolreprod.116.140277"},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Sato, T., Katagiri, K., Kojima, K., Komeya, M., Yao, M., and Ogawa, T. (2015). In Vitro Spermatogenesis in Explanted Adult Mouse Testis Tissues. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0130171"},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Matsumura, T., Sato, T., Abe, T., Sanjo, H., Katagiri, K., Kimura, H., Fujii, T., Tanaka, H., Hirabayashi, M., and Ogawa, T. (2021). Rat in vitro spermatogenesis promoted by chemical supplementations and oxygen-tension control. Sci. Rep., 11.","DOI":"10.1038\/s41598-021-82792-2"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"2098","DOI":"10.1038\/nprot.2013.138","article-title":"In vitro sperm production from mouse spermatogonial stem cell lines using an organ culture method","volume":"8","author":"Sato","year":"2013","journal-title":"Nat. Protoc."},{"key":"ref_63","doi-asserted-by":"crossref","unstructured":"Busada, J.T., and Geyer, C.B. (2016). The Role of Retinoic Acid (RA) in Spermatogonial Differentiation. Biol. Reprod., 94.","DOI":"10.1095\/biolreprod.115.135145"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.bbagrm.2014.05.019","article-title":"Role of retinoic acid receptor (RAR) signaling in post-natal male germ cell differentiation","volume":"1849","author":"Mark","year":"2015","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_65","doi-asserted-by":"crossref","unstructured":"Hogarth, C.A., Arnold, S., Kent, T., Mitchell, D., Isoherranen, N., and Griswold, M.D. (2015). Processive pulses of retinoic acid propel asynchronous and continuous murine sperm production. Biol. Reprod., 92.","DOI":"10.1095\/biolreprod.114.126326"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"15104","DOI":"10.1074\/jbc.M113.540211","article-title":"Inhibition of retinoic acid biosynthesis by the bisdichloroacetyldiamine WIN 18,446 markedly suppresses spermatogenesis and alters retinoid metabolism in mice","volume":"289","author":"Paik","year":"2014","journal-title":"J. Biol. Chem."},{"key":"ref_67","doi-asserted-by":"crossref","unstructured":"Sanjo, H., Komeya, M., Sato, T., Abe, T., Katagiri, K., Yamanaka, H., Ino, Y., Arakawa, N., Hirano, H., and Yao, T. (2018). In vitro mouse spermatogenesis with an organ culture method in chemically defined medium. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0192884"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"1","DOI":"10.4238\/gmr.15038602","article-title":"Isolation, proliferation, and induction of Bama mini-pig spermatogonial stem cells in vitro","volume":"15","author":"Zhao","year":"2016","journal-title":"Genet. Mol. Res."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1136\/jclinpath-2018-205579","article-title":"Investigation and management of subfertility","volume":"72","author":"Thurston","year":"2019","journal-title":"J. Clin. Pathol."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"258","DOI":"10.1016\/0012-1606(91)90468-I","article-title":"Human chorionic gonadotropin induces all stages of spermatogenesis in vitro in the male Japanese eel (Anguilla japonica)","volume":"146","author":"Miura","year":"1991","journal-title":"Dev. Biol."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"944","DOI":"10.1095\/biolreprod61.4.944","article-title":"Recombinant human insulin-like growth factor I stimulates all stages of 11-ketotestosterone-induced spermatogenesis in the Japanese eel, Anguilla japonica, in vitro","volume":"61","author":"Nader","year":"1999","journal-title":"Biol. Reprod."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"321","DOI":"10.2108\/zsj.19.321","article-title":"Comparative studies between in vivo and in vitro spermatogenesis of Japanese eel (Anguilla japonica)","volume":"19","author":"Miura","year":"2002","journal-title":"Zoolog Sci."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1111\/jpi.12327","article-title":"Melatonin promotes development of haploid germ cells from early developing spermatogenic cells of Suffolk sheep under in vitro condition","volume":"60","author":"Deng","year":"2016","journal-title":"J. Pineal Res."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1080\/19396368.2017.1358774","article-title":"In vitro effects of melatonin on colonization of neonate mouse spermatogonial stem cells","volume":"63","author":"Navid","year":"2017","journal-title":"Syst. Biol. Reprod. Med."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"122327","DOI":"10.1016\/j.jhazmat.2020.122327","article-title":"CdSe\/ZnS quantum dots induced spermatogenesis dysfunction via autophagy activation","volume":"398","author":"Yang","year":"2020","journal-title":"J. Hazard. Mater."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1006\/scdb.1998.0203","article-title":"The central role of Sertoli cells in spermatogenesis","volume":"9","author":"Griswold","year":"1998","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"dev200706","DOI":"10.1242\/dev.200706","article-title":"Sertoli cells are the source of stem cell factor for spermatogenesis","volume":"150","author":"Peng","year":"2023","journal-title":"Development"},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/j.theriogenology.2022.07.020","article-title":"Cryopreservation of testicular tissue from the dog (Canis familiaris) and wild boar (Sus scrofa) by slow freezing and vitrification: Differences in cryoresistance according to cell type","volume":"190","author":"Picazo","year":"2022","journal-title":"Theriogenology"},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1016\/j.cryobiol.2018.09.001","article-title":"Comparison of cryosurvival and spermatogenesis efficiency of cryopreserved neonatal mouse testicular tissue between three vitrification protocols and controlled-rate freezing","volume":"84","author":"Yildiz","year":"2018","journal-title":"Cryobiology"},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"744","DOI":"10.1093\/humupd\/dmw029","article-title":"Cryopreservation of testicular tissue or testicular cell suspensions: A pivotal step in fertility preservation","volume":"22","author":"Onofre","year":"2016","journal-title":"Hum. Reprod. Update"},{"key":"ref_81","doi-asserted-by":"crossref","unstructured":"Peris-Frau, P., Benito-Blanco, J., Martinez-Nevado, E., Toledano-Diaz, A., Castano, C., Velazquez, R., Pequeno, B., Martinez-Madrid, B., Esteso, M.C., and Santiago-Moreno, J. (2022). DNA integrity and viability of testicular cells from diverse wild species after slow freezing or vitrification. Front. Vet. Sci., 9.","DOI":"10.3389\/fvets.2022.1114695"},{"key":"ref_82","doi-asserted-by":"crossref","unstructured":"Silva, A.M., Pereira, A.G., Bezerra, L., Jeronimo, M.S., Pereira, A.F., Oliveira, M.F., Comizzoli, P., and Silva, A.R. (2022). Cryopreservation of Testicular Tissue from Adult Red-Rumped Agoutis (Dasyprocta leporina Linnaeus, 1758). Animals, 12.","DOI":"10.3390\/ani12060738"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/15\/8524\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T15:30:05Z","timestamp":1760110205000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/15\/8524"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,8,5]]},"references-count":82,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2024,8]]}},"alternative-id":["ijms25158524"],"URL":"https:\/\/doi.org\/10.3390\/ijms25158524","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,8,5]]}}}