Transcriptional Profiling of β-2MSPα-6THY1 Spermatogonial Stem Cells in Human Spermatogenesis

Overview

Journal Stem Cell Reports

Publisher Cell Press

Specialty Cell Biology

Date 2022 Mar 25

PMID 35334216

Authors

Maelle Givelet

Virginie Firlej

Bruno Lassalle

Anne Sophie Gille

Clementine Lapoujade

Isabelle Holtzman

Amandine Jarysta

Farahd Haghighirad

Florent Dumont

Sebastien Jacques

Franck Letourneur

Francoise Pflumio

Isabelle Allemand

Catherine Patrat

Nicolas Thiounn

Jean Philippe Wolf

Lydia Riou

Virginie Barraud-Lange

Pierre Fouchet

Affiliations

Soon will be listed here.

Abstract

Male infertility is responsible for approximately half of all cases of reproductive issues. Spermatogenesis originates in a small pool of spermatogonial stem cells (SSCs), which are of interest for therapy of infertility but remain not well defined in humans. Using multiparametric analysis of the side population (SP) phenotype and the α-6 integrin, THY1, and β-2 microglobulin cell markers, we identified a population of human primitive undifferentiated spermatogonia with the phenotype β-2 microglobulin (β-2M)SPα-6THY1, which is highly enriched in stem cells. By analyzing the expression signatures of this SSC-enriched population along with other germinal progenitors, we established an exhaustive transcriptome of human spermatogenesis. Transcriptome profiling of the human β-2MSPα-6THY1 population and comparison with the profile of mouse undifferentiated spermatogonia provide insights into the molecular networks and key transcriptional regulators regulating human SSCs, including the basic-helix-loop-helix (bHLH) transcriptional repressor HES1, which we show to be implicated in maintenance of SSCs in vitro.

Citing Articles

Characterisation and hierarchy of the spermatogonial stem cell compartment in human spermatogenesis by spectral cytometry using a 16-colors panel.

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Impact of the hypoxic microenvironment on spermatogonial stem cells in culture.

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Postnatal development of mouse spermatogonial stem cells as determined by immunophenotype, regenerative capacity, and long-term culture-initiating ability: a model for practical applications.

Song Y, Zhang X, Desmarais J, Nagano M Sci Rep. 2024; 14(1):2299.

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References

Nickkholgh B, Mizrak S, Korver C, van Daalen S, Meissner A, Repping S . Enrichment of spermatogonial stem cells from long-term cultured human testicular cells. Fertil Steril. 2014; 102(2):558-565.e5. DOI: 10.1016/j.fertnstert.2014.04.022. View

Wang P, McCarrey J, Yang F, Page D . An abundance of X-linked genes expressed in spermatogonia. Nat Genet. 2001; 27(4):422-6. DOI: 10.1038/86927. View

Zohni K, Zhang X, Tan S, Chan P, Nagano M . CD9 is expressed on human male germ cells that have a long-term repopulation potential after transplantation into mouse testes. Biol Reprod. 2012; 87(2):27. DOI: 10.1095/biolreprod.112.098913. View

Dovey S, Valli H, Hermann B, Sukhwani M, Donohue J, Castro C . Eliminating malignant contamination from therapeutic human spermatogonial stem cells. J Clin Invest. 2013; 123(4):1833-43. PMC: 3613920. DOI: 10.1172/JCI65822. View

Wang M, Liu X, Chang G, Chen Y, An G, Yan L . Single-Cell RNA Sequencing Analysis Reveals Sequential Cell Fate Transition during Human Spermatogenesis. Cell Stem Cell. 2018; 23(4):599-614.e4. DOI: 10.1016/j.stem.2018.08.007. View

Yu X, Alder J, Chun J, Friedman A, Heimfeld S, Cheng L . HES1 inhibits cycling of hematopoietic progenitor cells via DNA binding. Stem Cells. 2006; 24(4):876-88. DOI: 10.1634/stemcells.2005-0598. View

Kanatsu-Shinohara M, Tanaka T, Ogonuki N, Ogura A, Morimoto H, Cheng P . Myc/Mycn-mediated glycolysis enhances mouse spermatogonial stem cell self-renewal. Genes Dev. 2016; 30(23):2637-2648. PMC: 5204355. DOI: 10.1101/gad.287045.116. View

Medrano J, Rombaut C, Simon C, Pellicer A, Goossens E . Human spermatogonial stem cells display limited proliferation in vitro under mouse spermatogonial stem cell culture conditions. Fertil Steril. 2016; 106(6):1539-1549.e8. DOI: 10.1016/j.fertnstert.2016.07.1065. View

Hermann B, Cheng K, Singh A, Roa-de la Cruz L, Mutoji K, Chen I . The Mammalian Spermatogenesis Single-Cell Transcriptome, from Spermatogonial Stem Cells to Spermatids. Cell Rep. 2018; 25(6):1650-1667.e8. PMC: 6384825. DOI: 10.1016/j.celrep.2018.10.026. View

10.

Hasegawa K, Okamura Y, Saga Y . Notch signaling in Sertoli cells regulates cyclical gene expression of Hes1 but is dispensable for mouse spermatogenesis. Mol Cell Biol. 2011; 32(1):206-15. PMC: 3255714. DOI: 10.1128/MCB.06063-11. View

11.

Yeh J, Zhang X, Nagano M . Establishment of a short-term in vitro assay for mouse spermatogonial stem cells. Biol Reprod. 2007; 77(5):897-904. DOI: 10.1095/biolreprod.107.063057. View

12.

Kotzur T, Benavides-Garcia R, Mecklenburg J, Sanchez J, Reilly M, Hermann B . Granulocyte colony-stimulating factor (G-CSF) promotes spermatogenic regeneration from surviving spermatogonia after high-dose alkylating chemotherapy. Reprod Biol Endocrinol. 2017; 15(1):7. PMC: 5225630. DOI: 10.1186/s12958-016-0226-1. View

13.

Kageyama R, Ohtsuka T, Kobayashi T . Roles of Hes genes in neural development. Dev Growth Differ. 2008; 50 Suppl 1:S97-103. DOI: 10.1111/j.1440-169X.2008.00993.x. View

14.

de Rooij D . The nature and dynamics of spermatogonial stem cells. Development. 2017; 144(17):3022-3030. DOI: 10.1242/dev.146571. View

15.

Ehmcke J, Schlatt S . A revised model for spermatogonial expansion in man: lessons from non-human primates. Reproduction. 2006; 132(5):673-80. DOI: 10.1530/rep.1.01081. View

16.

Mevel R, Draper J, Lie-A-Ling M, Kouskoff V, Lacaud G . RUNX transcription factors: orchestrators of development. Development. 2019; 146(17). DOI: 10.1242/dev.148296. View

17.

Nakagawa T, Sharma M, Nabeshima Y, Braun R, Yoshida S . Functional hierarchy and reversibility within the murine spermatogenic stem cell compartment. Science. 2010; 328(5974):62-7. PMC: 2981100. DOI: 10.1126/science.1182868. View

18.

Falciatori I, Borsellino G, Haliassos N, Boitani C, Corallini S, Battistini L . Identification and enrichment of spermatogonial stem cells displaying side-population phenotype in immature mouse testis. FASEB J. 2003; 18(2):376-8. DOI: 10.1096/fj.03-0744fje. View

19.

Hermann B, Sukhwani M, Winkler F, Pascarella J, Peters K, Sheng Y . Spermatogonial stem cell transplantation into rhesus testes regenerates spermatogenesis producing functional sperm. Cell Stem Cell. 2012; 11(5):715-26. PMC: 3580057. DOI: 10.1016/j.stem.2012.07.017. View

20.

Zhou S, Schuetz J, Bunting K, Colapietro A, Sampath J, Morris J . The ABC transporter Bcrp1/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the side-population phenotype. Nat Med. 2001; 7(9):1028-34. DOI: 10.1038/nm0901-1028. View