{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T20:20:37Z","timestamp":1790022037388,"version":"4.0.1"},"reference-count":73,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2023,3,1]],"date-time":"2023-03-01T00:00:00Z","timestamp":1677628800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD)","award":["R01 HD090007"],"award-info":[{"award-number":["R01 HD090007"]}]},{"name":"Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD)","award":["U01 DA054170"],"award-info":[{"award-number":["U01 DA054170"]}]},{"name":"Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD)","award":["G12 MD007591"],"award-info":[{"award-number":["G12 MD007591"]}]},{"name":"Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD)","award":["DBI-1337513"],"award-info":[{"award-number":["DBI-1337513"]}]},{"name":"Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD)","award":["DBI-2018408"],"award-info":[{"award-number":["DBI-2018408"]}]},{"DOI":"10.13039\/100000026","name":"NIH BRAIN Initiative","doi-asserted-by":"publisher","award":["R01 HD090007"],"award-info":[{"award-number":["R01 HD090007"]}],"id":[{"id":"10.13039\/100000026","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000026","name":"NIH BRAIN Initiative","doi-asserted-by":"publisher","award":["U01 DA054170"],"award-info":[{"award-number":["U01 DA054170"]}],"id":[{"id":"10.13039\/100000026","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000026","name":"NIH BRAIN Initiative","doi-asserted-by":"publisher","award":["G12 MD007591"],"award-info":[{"award-number":["G12 MD007591"]}],"id":[{"id":"10.13039\/100000026","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000026","name":"NIH BRAIN Initiative","doi-asserted-by":"publisher","award":["DBI-1337513"],"award-info":[{"award-number":["DBI-1337513"]}],"id":[{"id":"10.13039\/100000026","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000026","name":"NIH BRAIN Initiative","doi-asserted-by":"publisher","award":["DBI-2018408"],"award-info":[{"award-number":["DBI-2018408"]}],"id":[{"id":"10.13039\/100000026","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"NIH","doi-asserted-by":"publisher","award":["R01 HD090007"],"award-info":[{"award-number":["R01 HD090007"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"NIH","doi-asserted-by":"publisher","award":["U01 DA054170"],"award-info":[{"award-number":["U01 DA054170"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"NIH","doi-asserted-by":"publisher","award":["G12 MD007591"],"award-info":[{"award-number":["G12 MD007591"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"NIH","doi-asserted-by":"publisher","award":["DBI-1337513"],"award-info":[{"award-number":["DBI-1337513"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"NIH","doi-asserted-by":"publisher","award":["DBI-2018408"],"award-info":[{"award-number":["DBI-2018408"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"name":"NSF","award":["R01 HD090007"],"award-info":[{"award-number":["R01 HD090007"]}]},{"name":"NSF","award":["U01 DA054170"],"award-info":[{"award-number":["U01 DA054170"]}]},{"name":"NSF","award":["G12 MD007591"],"award-info":[{"award-number":["G12 MD007591"]}]},{"name":"NSF","award":["DBI-1337513"],"award-info":[{"award-number":["DBI-1337513"]}]},{"name":"NSF","award":["DBI-2018408"],"award-info":[{"award-number":["DBI-2018408"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Antineoplastic treatments for cancer and other non-malignant disorders can result in long-term or permanent male infertility by ablating spermatogonial stem cells (SSCs). SSC transplantation using testicular tissue harvested before a sterilizing treatment is a promising approach for restoring male fertility in these cases, but a lack of exclusive biomarkers to unequivocally identify prepubertal SSCs limits their therapeutic potential. To address this, we performed single-cell RNA-seq on testis cells from immature baboons and macaques and compared these cells with published data from prepubertal human testis cells and functionally-defined mouse SSCs. While we found discrete groups of human spermatogonia, baboon and rhesus spermatogonia appeared less heterogenous. A cross-species analysis revealed cell types analogous to human SSCs in baboon and rhesus germ cells, but a comparison with mouse SSCs revealed significant differences with primate SSCs. Primate-specific SSC genes were enriched for components and regulators of the actin cytoskeleton and participate in cell-adhesion, which may explain why the culture conditions for rodent SSCs are not appropriate for primate SSCs. Furthermore, correlating the molecular definitions of human SSC, progenitor and differentiating spermatogonia with the histological definitions of Adark\/Apale spermatogonia indicates that both SSCs and progenitor spermatogonia are Adark, while Apale spermatogonia appear biased towards differentiation. These results resolve the molecular identity of prepubertal human SSCs, define novel pathways that could be leveraged for advancing their selection and propagation in vitro, and confirm that the human SSC pool resides entirely within Adark spermatogonia.<\/jats:p>","DOI":"10.3390\/ijms24054755","type":"journal-article","created":{"date-parts":[[2023,3,2]],"date-time":"2023-03-02T03:29:00Z","timestamp":1677727740000},"page":"4755","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Conserved Transcriptome Features Define Prepubertal Primate Spermatogonial Stem Cells as Adark Spermatogonia and Identify Unique Regulators"],"prefix":"10.3390","volume":"24","author":[{"given":"Anukriti","family":"Singh","sequence":"first","affiliation":[{"name":"Department of Neuroscience, Developmental and Regenerative Biology, The University of Texas at San Antonio, 1 UTSA Circle, San Antonio, TX 78249, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0503-3616","authenticated-orcid":false,"given":"Brian P.","family":"Hermann","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Developmental and Regenerative Biology, The University of Texas at San Antonio, 1 UTSA Circle, San Antonio, TX 78249, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,3,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Lambertini, M., Del Mastro, L., Pescio, M.C., Andersen, C.Y., Azim, H.A., Peccatori, F.A., Costa, M., Revelli, A., Salvagno, F., and Gennari, A. (2016). Cancer and fertility preservation: International recommendations from an expert meeting. BMC Med., 14.","DOI":"10.1186\/s12916-015-0545-7"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1093\/humrep\/12.1.87","article-title":"Full-term delivery following intracytoplasmic sperm injection with spermatozoa extracted from frozen-thawed testicular tissue","volume":"12","author":"Khalifeh","year":"1997","journal-title":"Hum. Reprod."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3256","DOI":"10.12659\/MSM.913513","article-title":"Sperm Cryopreservation for Male Cancer Patients: More than 10 Years of Experience, in Beijing China","volume":"25","author":"Fu","year":"2019","journal-title":"Med. Sci. Monit."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/0140-6736(92)92425-F","article-title":"Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte","volume":"340","author":"Palermo","year":"1992","journal-title":"Lancet"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1136\/medethics-2016-104042","article-title":"Ethics of fertility preservation for prepubertal children: Should clinicians offer procedures where efficacy is largely unproven?","volume":"44","author":"McDougall","year":"2018","journal-title":"J. Med. Ethics"},{"key":"ref_6","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_7","doi-asserted-by":"crossref","first-page":"1677","DOI":"10.1093\/humrep\/dey240","article-title":"Spermatogonial quantity in human prepubertal testicular tissue collected for fertility preservation prior to potentially sterilizing therapy","volume":"33","author":"Stukenborg","year":"2018","journal-title":"Hum. Reprod."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"966","DOI":"10.1093\/humrep\/dez043","article-title":"Testicular tissue cryopreservation: 8 years of experience from a coordinated network of academic centers","volume":"34","author":"Peters","year":"2019","journal-title":"Hum. Reprod."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1249","DOI":"10.1093\/humrep\/des077","article-title":"Fertile offspring derived from mouse spermatogonial stem cells cryopreserved for more than 14 years","volume":"27","author":"Wu","year":"2012","journal-title":"Hum. Reprod."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1314","DOI":"10.1126\/science.aav2914","article-title":"Autologous grafting of cryopreserved prepubertal rhesus testis produces sperm and offspring","volume":"363","author":"Fayomi","year":"2019","journal-title":"Science"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2127","DOI":"10.1001\/jama.2009.1689","article-title":"Propagation of Human Spermatogonial Stem Cells In Vitro","volume":"302","author":"Mizrak","year":"2009","journal-title":"JAMA"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"21672","DOI":"10.1073\/pnas.0912432106","article-title":"Prepubertal human spermatogonia and mouse gonocytes share conserved gene expression of germline stem cell regulatory molecules","volume":"106","author":"Wu","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2416","DOI":"10.1001\/jama.2011.791","article-title":"In Vitro Propagation of Human Prepubertal Spermatogonial Stem Cells","volume":"305","author":"Akhondi","year":"2011","journal-title":"JAMA"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1038\/aja.2009.21","article-title":"Xeno-free culture of human spermatogonial stem cells supported by human embryonic stem cell-derived fibroblast-like cells","volume":"11","author":"Chen","year":"2009","journal-title":"Asian J. Androl."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1111\/j.1439-0272.2010.01135.x","article-title":"Effects of basic fibroblast growth factor and leukaemia inhibitory factor on proliferation and short-term culture of human spermatogonial stem cells","volume":"44","author":"Mirzapour","year":"2012","journal-title":"Andrologia"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"957","DOI":"10.1007\/s10815-012-9817-8","article-title":"Proliferation of small number of human spermatogonial stem cells obtained from azoospermic patients","volume":"29","author":"Koruji","year":"2012","journal-title":"J. Assist. Reprod. Genet."},{"key":"ref_17","first-page":"54","article-title":"Isolation and Culture of Human Spermatogonial Stem Cells Derived from Testis Biopsy","volume":"5","author":"Goharbakhsh","year":"2013","journal-title":"Avicenna J. Med. Biotechnol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1112","DOI":"10.1177\/1535370215590822","article-title":"Expansion and long-term culture of human spermatogonial stem cells via the activation of SMAD3 and AKT pathways","volume":"240","author":"Guo","year":"2015","journal-title":"Exp. Biol. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1002\/jcp.21789","article-title":"Asymmetric Distribution of UCH-L1 in Spermatogonia Is Associated with Maintenance and Differentiation of Spermatogonial Stem Cells","volume":"220","author":"Luo","year":"2009","journal-title":"J. Cell. Physiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1002\/aja.1001420203","article-title":"Spermatogonial stem cells in the albino rat","volume":"142","author":"Clermont","year":"1975","journal-title":"Am. J. Anat."},{"key":"ref_21","first-page":"135","article-title":"Renewal of type A spermatogonia in adult rats","volume":"16","author":"Bartmanska","year":"1983","journal-title":"Cell Tissue Kinet."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1002\/aja.1001220205","article-title":"Re-examination of spermatogonial renewal in the rat by means of seminiferous tubules mounted \u201cin toto\u201d","volume":"122","author":"Clermont","year":"1968","journal-title":"Am. J. Anat."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1152\/physrev.1972.52.1.198","article-title":"Kinetics of spermatogenesis in mammals: Seminiferous epithelium cycle and spermatogonial renewal","volume":"52","author":"Clermont","year":"1972","journal-title":"Physiol. Rev."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1002\/aja.1001120103","article-title":"The cycle of the seminiferous epithelium in man","volume":"112","author":"Clermont","year":"1963","journal-title":"Am. J. Anat."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1002\/ar.1091690305","article-title":"Spermatogonial stem-cell renewal in the mouse","volume":"169","author":"Oakberg","year":"1971","journal-title":"Anat. Rec."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"R55","DOI":"10.1530\/REP-17-0034","article-title":"A revised Asingle model to explain stem cell dynamics in the mouse male germline","volume":"154","author":"Lord","year":"2017","journal-title":"Reproduction"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1126\/science.1182868","article-title":"Functional Hierarchy and Reversibility Within the Murine Spermatogenic Stem Cell Compartment","volume":"328","author":"Nakagawa","year":"2010","journal-title":"Science"},{"key":"ref_28","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_29","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1002\/ar.1091690306","article-title":"The spermatogonial stem cell population in adult rats. I. Their morphology, proliferation and maturation","volume":"169","author":"Huckins","year":"1971","journal-title":"Anat. Rec."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/0027-5107(93)90159-D","article-title":"A quantitative study of spermatogonial multiplication and stem cell renewal in the C3H\/101 F1 hybrid mouse","volume":"290","author":"Tagelenbosch","year":"1993","journal-title":"Mutat. Res. Mol. Mech. Mutagen."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1095\/biolreprod.103.016352","article-title":"Homing Efficiency and Proliferation Kinetics of Male Germ Line Stem Cells Following Transplantation in Mice","volume":"69","author":"Nagano","year":"2003","journal-title":"Biol. Reprod."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1351","DOI":"10.1101\/gad.240465.114","article-title":"Functional and molecular features of the ID4+ germline stem cell population in mouse testes","volume":"28","author":"Oatley","year":"2014","journal-title":"Genes Dev."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1650","DOI":"10.1016\/j.celrep.2018.10.026","article-title":"The Mammalian Spermatogenesis Single-Cell Transcriptome, from Spermatogonial Stem Cells to Spermatids","volume":"25","author":"Hermann","year":"2018","journal-title":"Cell Rep."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"624","DOI":"10.1242\/dev.146928","article-title":"ID4 levels dictate the stem cell state in mouse spermatogonia","volume":"144","author":"Helsel","year":"2017","journal-title":"Development"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1002\/aja.1001180211","article-title":"Renewal of spermatogonia in man","volume":"118","author":"Clermont","year":"1966","journal-title":"Am. J. Anat."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1002\/aja.1001040204","article-title":"Differentiation and renewal of spermatogonia in the monkey, Macacus rhesus","volume":"104","author":"Clermont","year":"1959","journal-title":"Am. J. Anat."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1002\/aja.1001260106","article-title":"Two classes of spermatogonial stem cells in the monkey (Cercopithecus aethiops)","volume":"126","author":"Clermont","year":"1969","journal-title":"Am. J. Anat."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"705","DOI":"10.1016\/S0015-0282(16)36120-9","article-title":"Spermatogenesis in man. A study of the spermatogonial population","volume":"17","author":"Clermont","year":"1966","journal-title":"Fertil. Steril."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1093\/humupd\/dmk001","article-title":"Spermatogonial stem cells: Questions, models and perspectives","volume":"12","author":"Ehmcke","year":"2006","journal-title":"Hum. Reprod. Update"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1095\/biolreprod35.1.199","article-title":"Renewal of Spermatogonia in the Monkey (Macaca Fascilularis)","volume":"35","author":"Fouquet","year":"1986","journal-title":"Biol. Reprod."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1530\/REP-09-0255","article-title":"Spermatogonial stem cells in higher primates: Are there differences from those in rodents?","volume":"139","author":"Hermann","year":"2010","journal-title":"Reproduction"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1501","DOI":"10.1016\/j.celrep.2019.01.045","article-title":"The Neonatal and Adult Human Testis Defined at the Single-Cell Level","volume":"26","author":"Sohni","year":"2019","journal-title":"Cell Rep."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"262","DOI":"10.1016\/j.stem.2019.12.005","article-title":"The Dynamic Transcriptional Cell Atlas of Testis Development during Human Puberty","volume":"26","author":"Guo","year":"2020","journal-title":"Cell Stem Cell"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1038\/s41422-018-0099-2","article-title":"The adult human testis transcriptional cell atlas","volume":"28","author":"Guo","year":"2018","journal-title":"Cell Res."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"5683","DOI":"10.1038\/s41467-020-19414-4","article-title":"Single-cell analysis of developing and azoospermia human testicles reveals central role of Sertoli cells","volume":"11","author":"Zhao","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1016\/j.stem.2018.08.007","article-title":"Single-Cell RNA Sequencing Analysis Reveals Sequential Cell Fate Transition during Human Spermatogenesis","volume":"23","author":"Wang","year":"2018","journal-title":"Cell Stem Cell"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"764","DOI":"10.1016\/j.stem.2020.12.004","article-title":"Single-cell analysis of the developing human testis reveals somatic niche cell specification and fetal germline stem cell establishment","volume":"28","author":"Guo","year":"2021","journal-title":"Cell Stem Cell"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"2095","DOI":"10.1093\/humrep\/deac157","article-title":"Metabolic transitions define spermatogonial stem cell maturation","volume":"37","author":"Voigt","year":"2022","journal-title":"Hum. Reprod."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"100395","DOI":"10.1016\/j.xcrm.2021.100395","article-title":"Single-cell RNA-seq unravels alterations of the human spermatogonial stem cell compartment in patients with impaired spermatogenesis","volume":"2","author":"Tekath","year":"2021","journal-title":"Cell Rep. Med."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1093\/biolre\/ioz088","article-title":"What has single-cell RNA-seq taught us about mammalian spermatogenesis?","volume":"101","author":"Suzuki","year":"2019","journal-title":"Biol. Reprod."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Robertson, M.J., Kent, K., Tharp, N., Nozawa, K., Dean, L., Mathew, M., Grimm, S.L., Yu, Z., L\u00e9gar\u00e9, C., and Fujihara, Y. (2020). Large-scale discovery of male reproductive tract-specific genes through analysis of RNA-seq datasets. BMC Biol., 18.","DOI":"10.1186\/s12915-020-00826-z"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1530\/REP-11-0290","article-title":"Differential marker protein expression specifies rarefaction zone-containing human Adark spermatogonia","volume":"143","author":"Staege","year":"2012","journal-title":"Reproduction"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1007\/s00418-012-0991-7","article-title":"Fibroblast growth factor receptor 3 is highly expressed in rarely dividing human type A spermatogonia","volume":"138","author":"Staege","year":"2012","journal-title":"Histochem. Cell Biol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1244","DOI":"10.1016\/j.fertnstert.2015.07.1134","article-title":"Cryopreservation of testicular tissue before long-term testicular cell culture does not alter in vitro cell dynamics","volume":"104","author":"Baert","year":"2015","journal-title":"Fertil. Steril."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"4778","DOI":"10.1128\/MCB.23.14.4778-4787.2003","article-title":"Mek2 Is Dispensable for Mouse Growth and Development","volume":"23","author":"Roy","year":"2003","journal-title":"Mol. Cell. Biol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"463","DOI":"10.1016\/j.stem.2011.08.011","article-title":"Rac Mediates Mouse Spermatogonial Stem Cell Homing to Germline Niches by Regulating Transmigration through the Blood-Testis Barrier","volume":"9","author":"Takashima","year":"2011","journal-title":"Cell Stem Cell"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"8321","DOI":"10.1074\/jbc.M105363200","article-title":"The Small GTPase Rac3 Interacts with the Integrin-binding Protein CIB and Promotes Integrin \u03b1IIb\u03b23-mediated Adhesion and Spreading","volume":"277","author":"Haataja","year":"2002","journal-title":"J. Biol. Chem."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/S0091-679X(08)00004-6","article-title":"Culture of Rodent Spermatogonial Stem Cells, Male Germline Stem Cells of the Postnatal Animal","volume":"86","author":"Kubota","year":"2008","journal-title":"Methods Cell Biol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1242\/dev.125.4.679","article-title":"Nanos and Pumilio have critical roles in the development and function of Drosophila germline stem cells","volume":"125","author":"Forbes","year":"1998","journal-title":"Development"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/j.mod.2010.12.001","article-title":"Nanos1 functions as a translational repressor in the Xenopus germline","volume":"128","author":"Lai","year":"2011","journal-title":"Mech. Dev."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"4861","DOI":"10.1242\/dev.126.21.4861","article-title":"Nos-1 and nos-2, two genes related to Drosophila nanos, regulate primordial germ cell development and survival in Caenorhabditis elegans","volume":"126","author":"Subramaniam","year":"1999","journal-title":"Development"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1242\/dev.02697","article-title":"Functional redundancy among Nanos proteins and a distinct role of Nanos2 during male germ cell development","volume":"134","author":"Suzuki","year":"2007","journal-title":"Development"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"1239","DOI":"10.1126\/science.1085222","article-title":"Conserved Role of nanos Proteins in Germ Cell Development","volume":"301","author":"Tsuda","year":"2003","journal-title":"Science"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"1663","DOI":"10.1098\/rstb.2010.0026","article-title":"Spermatogonial stem cell regulation and spermatogenesis","volume":"365","author":"Phillips","year":"2010","journal-title":"Philos. Trans. R. Soc. B Biol. Sci."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"3022","DOI":"10.1242\/dev.146571","article-title":"The nature and dynamics of spermatogonial stem cells","volume":"144","year":"2017","journal-title":"Development"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"109915","DOI":"10.1016\/j.celrep.2021.109915","article-title":"Dissecting mammalian spermatogenesis using spatial transcriptomics","volume":"37","author":"Chen","year":"2021","journal-title":"Cell Rep."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"2330","DOI":"10.1634\/stemcells.2007-0143","article-title":"Characterization, Cryopreservation, and Ablation of Spermatogonial Stem Cells in Adult Rhesus Macaques","volume":"25","author":"Hermann","year":"2007","journal-title":"Stem Cells"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"14049","DOI":"10.1038\/ncomms14049","article-title":"Massively parallel digital transcriptional profiling of single cells","volume":"8","author":"Zheng","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"1888","DOI":"10.1016\/j.cell.2019.05.031","article-title":"Comprehensive Integration of Single-Cell Data","volume":"177","author":"Stuart","year":"2019","journal-title":"Cell"},{"key":"ref_70","unstructured":"Wickham, H., and Henry, L. (2022, August 11). Tidyr: Tidy Messy Data. R Package Version 1.0.0. Available online: https:\/\/cran.r-project.org\/package=tidyr."},{"key":"ref_71","first-page":"1","article-title":"Ggplot2: Elegant graphics for data analysis","volume":"35","year":"2010","journal-title":"J. Stat. Softw."},{"key":"ref_72","doi-asserted-by":"crossref","unstructured":"Chen, E.Y., Tan, C.M., Kou, Y., Duan, Q., Wang, Z., Meirelles, G.V., Clark, N.R., and Ma\u2019Ayan, A. (2013). Enrichr: Interactive and collaborative HTML5 gene list enrichment analysis tool. BMC Bioinform., 14.","DOI":"10.1186\/1471-2105-14-128"},{"key":"ref_73","doi-asserted-by":"crossref","unstructured":"Pico, A., Kelder, T., van Iersel, M., Hanspers, K., Conklin, B.R., and Evelo, C. (2008). WikiPathways: Pathway Editing for the People. PLoS Biol., 6.","DOI":"10.1371\/journal.pbio.0060184"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/5\/4755\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:45:18Z","timestamp":1760121918000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/5\/4755"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,3,1]]},"references-count":73,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2023,3]]}},"alternative-id":["ijms24054755"],"URL":"https:\/\/doi.org\/10.3390\/ijms24054755","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,3,1]]}}}