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Being critical to male reproduction, it is generally assumed that spermatogenesis starts and ends in equivalent transcriptional states in related species. Based on single-cell gene expression profiling, it has been proposed that undifferentiated human spermatogonia can be subclassified into four heterogenous subtypes, termed states 0, 0A, 0B, and 1. To increase the resolution of the undifferentiated compartment and trace the origin of the spermatogenic trajectory, we re-analysed the single-cell (sc) RNA-sequencing libraries of 34 post-pubescent human testes to generate an integrated atlas of germ cell differentiation. We then used this atlas to perform comparative analyses of the putative SSC transcriptome both across human development (using 28 foetal and pre-pubertal scRNA-seq libraries) and across species (including data from sheep, pig, buffalo, rhesus and cynomolgus macaque, rat, and mouse). Alongside its detailed characterisation, we show that the transcriptional heterogeneity of the undifferentiated spermatogonial cell compartment varies not only between species but across development. Our findings associate \u2018state 0B\u2019 with a suppressive transcriptomic programme that, in adult humans, acts to functionally oppose proliferation and maintain cells in a ready-to-react state. Consistent with this conclusion, we show that human foetal germ cells\u2014which are mitotically arrested\u2014can be characterised solely as state 0B. While germ cells with a state 0B signature are also present in foetal mice (and are likely conserved at this stage throughout mammals), they are not maintained into adulthood. We conjecture that in rodents, the foetal-like state 0B differentiates at birth into the renewing SSC population, whereas in humans it is maintained as a reserve population, supporting testicular homeostasis over a longer reproductive lifespan while reducing mutagenic load. Together, these results suggest that SSCs adopt differing evolutionary strategies across species to ensure fertility and genome integrity over vastly differing life histories and reproductive timeframes.<\/jats:p>","DOI":"10.3390\/cells13090742","type":"journal-article","created":{"date-parts":[[2024,4,24]],"date-time":"2024-04-24T10:18:36Z","timestamp":1713953916000},"page":"742","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Adult Human, but Not Rodent, Spermatogonial Stem Cells Retain States with a Foetal-like Signature"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9341-2562","authenticated-orcid":false,"given":"Stephen J.","family":"Bush","sequence":"first","affiliation":[{"name":"MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford OX3 9DS, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-2571-9140","authenticated-orcid":false,"given":"Rafail","family":"Nikola","sequence":"additional","affiliation":[{"name":"MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford OX3 9DS, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seungmin","family":"Han","sequence":"additional","affiliation":[{"name":"Wellcome Trust\/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge CB2 1QN, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0241-1455","authenticated-orcid":false,"given":"Shinnosuke","family":"Suzuki","sequence":"additional","affiliation":[{"name":"Division of Germ Cell Biology, National Institute for Basic Biology, National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki 444-8787, Japan"},{"name":"Graduate Institute for Advanced Studies, SOKENDAI, 5-1 Higashiyama, Myodaiji, Okazaki 444-8787, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shosei","family":"Yoshida","sequence":"additional","affiliation":[{"name":"Division of Germ Cell Biology, National Institute for Basic Biology, National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki 444-8787, Japan"},{"name":"Graduate Institute for Advanced Studies, SOKENDAI, 5-1 Higashiyama, Myodaiji, Okazaki 444-8787, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Benjamin D.","family":"Simons","sequence":"additional","affiliation":[{"name":"Wellcome Trust\/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge CB2 1QN, UK"},{"name":"Wellcome\u2014MRC Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge CB2 0AW, UK"},{"name":"Department of Applied Mathematics and Theoretical Physics, Centre for Mathematical Science, University of Cambridge, Cambridge CB3 0WA, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anne","family":"Goriely","sequence":"additional","affiliation":[{"name":"MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford OX3 9DS, UK"},{"name":"NIHR Biomedical Research Centre, Oxford OX3 7JX, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,4,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1038\/s41586-023-05752-y","article-title":"Evolution of the germline mutation rate across vertebrates","volume":"615","author":"Bergeron","year":"2023","journal-title":"Nature"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"672","DOI":"10.1038\/s41576-021-00376-2","article-title":"The origin of human mutation in light of genomic data","volume":"22","author":"Seplyarskiy","year":"2021","journal-title":"Nat. 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