{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T03:16:37Z","timestamp":1790738197087,"version":"4.1.0"},"reference-count":62,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,3,17]],"date-time":"2023-03-17T00:00:00Z","timestamp":1679011200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001665","name":"Agence Nationale de la Recherche","doi-asserted-by":"publisher","award":["ANR-21-CE14-0068"],"award-info":[{"award-number":["ANR-21-CE14-0068"]}],"id":[{"id":"10.13039\/501100001665","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Endocrinol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Suitable cryopreservation procedures of pre-pubertal testicular tissue associated with efficient culture conditions are crucial in the fields of fertility preservation and restoration. <jats:italic>In vitro<\/jats:italic> spermatogenesis remains a challenging technical procedure to undergo a complete spermatogenesis.The number of haploid cells and more specifically the spermatic yield produced <jats:italic>in vitro<\/jats:italic> in mice is still extremely low compared to age-matched <jats:italic>in vivo<\/jats:italic> controls and this procedure has never yet been successfully transferred to humans.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>To evaluate the impact of in vitro culture and freezing procedure, pre-pubertal testicular mice testes were directly cultured until day 4 (D4), D16 and D30 or cryopreserved by controlled slow freezing then cultured until D30. Testes composed of a panel of 6.5 dpp (days postpartum), 10.5 dpp, 22.5 dpp, and 36.5 dpp mice were used as <jats:italic>in vivo<\/jats:italic> controls. Testicular tissues were assessed by histological (HES) and immunofluorescence (stimulated by retinoic acid gene 8, STRA8) analyses. Moreover, a detailed transcriptome evaluation study has been carried out to study the gene expression patterns throughout the first <jats:italic>in vitro<\/jats:italic> spermatogenic wave.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Transcriptomic analyses reveal that cultured tissues expression profiles are almost comparable between D16 and D30; highlighting an abnormal kinetic throughout the second half of the first spermatogenesis during <jats:italic>in vitro<\/jats:italic> cultures. In addition, testicular explants have shown dysregulation of their transcriptomic profile compared to controls with genes related to inflammation response, insulin-like growth factor and genes involved in steroidogenesis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>The present work first shows that cryopreservation had very little impact on gene expression in testicular tissue, either directly after thawing or after 30 days in culture. Transcriptomic analysis of testis tissue samples is highly informative due to the large number of expressed genes and identified isoforms. This study provides a very valuable basis for future studies concerning <jats:italic>in vitro<\/jats:italic> spermatogenesis in mice.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fendo.2023.1112834","type":"journal-article","created":{"date-parts":[[2023,3,17]],"date-time":"2023-03-17T05:17:01Z","timestamp":1679030221000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":15,"title":["Throughout in vitro first spermatogenic wave: Next-generation sequencing gene expression patterns of fresh and cryopreserved prepubertal mice testicular tissue explants"],"prefix":"10.3389","volume":"14","author":[{"given":"Ludovic","family":"Dumont","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"H\u00e9l\u00e8ne","family":"Lopez Maestre","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fr\u00e9d\u00e9ric","family":"Chalmel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Louise","family":"Huber","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aur\u00e9lie","family":"Rives-Feraille","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laura","family":"Moutard","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fr\u00e9d\u00e9rique","family":"Bateux","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christine","family":"Rondanino","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nathalie","family":"Rives","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,3,17]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1159\/000078189","article-title":"Role of retinoid signaling in the regulation of spermatogenesis","volume":"105","author":"Chung","year":"2004","journal-title":"Cytogenet Genome Res"},{"key":"B2","doi-asserted-by":"publisher","first-page":"83e83","DOI":"10.1111\/j.1365-2605.1993.tb01156.x","article-title":"Histological and histopathological evaluation of the testis","volume":"16","author":"Russell","year":"1993","journal-title":"Int J Androl."},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1002\/j.1939-4640.1986.tb00924.x","article-title":"The effect of selective destruction and regeneration of rat leydig cells on the intratesticular distribution of testosterone and morphology of the seminiferous epithelium","volume":"7","author":"Bartlett","year":"1986","journal-title":"J Androl."},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1093\/biolre\/ioy059","article-title":"Leydig cells: formation, function, and regulation","volume":"99","author":"Zirkin","year":"2018","journal-title":"Biol Reprod"},{"key":"B5","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1111\/andr.12165","article-title":"The sertoli cell: one hundred fifty years of beauty and plasticity","volume":"4","author":"Fran\u00e7a","year":"2016","journal-title":"Andrology."},{"key":"B6","doi-asserted-by":"publisher","first-page":"87","DOI":"10.1093\/biolre\/ioy027","article-title":"50 years of spermatogenesis: Sertoli cells and their interactions with germ cells","volume":"99","author":"Griswold","year":"2018","journal-title":"Biol Reprod"},{"key":"B7","doi-asserted-by":"publisher","first-page":"779","DOI":"10.1016\/0960-0760(92)90307-5","article-title":"Steroidogenic enzymes: structure, function, and role in regulation of steroid hormone biosynthesis","volume":"43","author":"Hanukoglu","year":"1992","journal-title":"J Steroid Biochem Mol Biol"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod.103.026880","article-title":"The murine testicular transcriptome: Characterizing gene expression in the testis during the progression of spermatogenesis","volume":"71","author":"Shima","year":"2004","journal-title":"Biol Reprod"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1093\/humrep\/dev190","article-title":"A European perspective on testicular tissue cryopreservation for fertility preservation in prepubertal and adolescent boys","volume":"30","author":"Picton","year":"2015","journal-title":"Hum Reprod"},{"key":"B10","doi-asserted-by":"publisher","first-page":"1454","DOI":"10.3390\/ijms21041454","article-title":"Exposure to chemotherapy during childhood or adulthood and consequences on spermatogenesis and Male fertility","volume":"21","author":"Delessard","year":"2020","journal-title":"Int J Mol Sci"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1093\/humupd\/dmp054","article-title":"Options for fertility preservation in prepubertal boys","volume":"16","author":"Wyns","year":"2010","journal-title":"Hum Reprod Update."},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1530\/REP-14-0249","article-title":"Xenografting of testicular tissue pieces: 12 years of an in vivo spermatogenesis system","volume":"148","author":"Arregui","year":"2014","journal-title":"Reproduction."},{"key":"B13","doi-asserted-by":"publisher","first-page":"2605","DOI":"10.1038\/s41598-017-02604-4","article-title":"Dysregulation of angiogenesis-specific signalling in adult testis results in xenograft degeneration","volume":"7","author":"Devi","year":"2017","journal-title":"Sci Rep"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1093\/humrep\/dem085","article-title":"Xenotransplantation of testicular tissue into nude mice can be used for detecting leukemic cell contamination","volume":"22","author":"Hou","year":"2007","journal-title":"Hum Reprod"},{"key":"B15","doi-asserted-by":"publisher","first-page":"472","DOI":"10.1038\/ncomms1478","article-title":"In vitro production of fertile sperm from murine spermatogonial stem cell lines","volume":"2","author":"Sato","year":"2011","journal-title":"Nat Commun"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0116660","article-title":"Retinol improves in vitro differentiation of pre-pubertal mouse spermatogonial stem cells into sperm during the first wave of spermatogenesis","volume":"10","author":"Arkoun","year":"2015","journal-title":"PLoS One"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1111\/andr.12042","article-title":"Assessment of the optimal vitrification protocol for pre-pubertal mice testes leading to successful in vitro production of flagellated spermatozoa","volume":"3","author":"Dumont","year":"2015","journal-title":"Andrology."},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1007\/s00441-015-2341-2","article-title":"Does soaking temperature during controlled slow freezing of pre-pubertal mouse testes influence course of in vitro spermatogenesis","volume":"364","author":"Arkoun","year":"2016","journal-title":"Cell Tissue Res"},{"key":"B19","doi-asserted-by":"publisher","first-page":"4320","DOI":"10.1038\/ncomms5320","article-title":"Offspring production with sperm grown in vitro from cryopreserved testis tissues","volume":"5","author":"Yokonishi","year":"2014","journal-title":"Nat Commun"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0061558","article-title":"Transcriptome profiling of the murine testis during the first wave of spermatogenesis","volume":"8","author":"Laiho","year":"2013","journal-title":"PLoS One"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2020.06.161","article-title":"Transcriptome analysis reveals inadequate spermatogenesis and immediate radical immune reactions during organ culture in vitro spermatogenesis","volume":"530","author":"Abe","year":"2020","journal-title":"Biochem Biophys Res Commun"},{"key":"B22","doi-asserted-by":"publisher","first-page":"106482","DOI":"10.1016\/j.dib.2020.106482","article-title":"Time-course microarray transcriptome data of in vitro cultured testes and age-matched in vivo testes","author":"Abe","year":"2020","journal-title":"Data Brief."},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-416022-4.00009-3","article-title":"Spermatogonial stem cell functions in physiological and pathological conditions","volume":"107","author":"Yang","year":"2014","journal-title":"Curr Top Dev Biol"},{"key":"B24","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1093\/humrep\/dem355","article-title":"Comparison of conditions for cryopreservation of testicular tissue from immature mice","volume":"23","author":"Milazzo","year":"2008","journal-title":"Hum Reprod"},{"key":"B25","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1093\/bioinformatics\/bts635","article-title":"STAR: ultrafast universal RNA-seq aligner","volume":"29","author":"Dobin","year":"2013","journal-title":"Bioinformatics."},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1101\/gr.133009.111","article-title":"Systematic identification of long noncoding RNAs expressed during zebrafish embryogenesis","volume":"22","author":"Pauli","year":"2012","journal-title":"Genome Res"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2012.016","article-title":"Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and cufflinks","volume":"7","author":"Trapnell","year":"2012","journal-title":"Nat Protoc"},{"key":"B28","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1095\/biolreprod.114.118166","article-title":"High-resolution profiling of novel transcribed regions during rat spermatogenesis","volume":"91","author":"Chalmel","year":"2014","journal-title":"Biol Reprod"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1210\/me.2014-1356","article-title":"Research resource: the dynamic transcriptional profile of sertoli cells during the progression of spermatogenesis","volume":"29","author":"Zimmermann","year":"2015","journal-title":"Mol Endocrinol"},{"key":"B30","doi-asserted-by":"publisher","first-page":"5893","DOI":"10.3390\/ijms23115893","article-title":"Understanding the underlying molecular mechanisms of meiotic arrest during In vitro spermatogenesis in rat prepubertal testicular tissue","volume":"23","author":"Saulnier","year":"2022","journal-title":"Int J Mol Sci"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2016.095","article-title":"Transcript-level expression analysis of RNA-seq experiments with HISAT, StringTie and ballgown","volume":"11","author":"Pertea","year":"2016","journal-title":"Nat Protoc"},{"key":"B32","doi-asserted-by":"publisher","first-page":"86","DOI":"10.1186\/1471-2105-9-86","article-title":"The annotation, mapping, expression and network (AMEN) suite of tools for molecular systems biology","volume":"9","author":"Chalmel","year":"2008","journal-title":"BMC Bioinf"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/bth449","article-title":"limmaGUI: a graphical user interface for linear modeling of microarray data","volume":"20","author":"Wettenhall","year":"2004","journal-title":"Bioinformatics."},{"key":"B34","doi-asserted-by":"publisher","first-page":"1","DOI":"10.18637\/jss.v025.i01","article-title":"FactoMineR: An r package for multivariate analysis","volume":"25","author":"L\u00ea","year":"2008","journal-title":"J Stat Software"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btu638","article-title":"HTSeq\u2013a Python framework to work with high-throughput sequencing data","volume":"31","author":"Anders","year":"2015","journal-title":"Bioinformatics"},{"key":"B36","doi-asserted-by":"publisher","first-page":"289","DOI":"10.1111\/j.2517-6161.1995.tb02031.x","article-title":"Controlling the false discovery rate: a practical and powerful approach to multiple testing","volume":"57","author":"Benjamini","year":"1995","journal-title":"J R Stat Soc Ser B"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1093\/molehr\/gax054","article-title":"Evaluation of apoptotic- and autophagic-related protein expressions before and after IVM of fresh, slow-frozen and vitrified pre-pubertal mouse testicular tissue","volume":"23","author":"Dumont","year":"2017","journal-title":"Mol Hum Reprod"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1007\/s10439-020-02535-8","article-title":"Improving freezing protocols and organotypic culture: A histological study on rat prepubertal testicular tissue","volume":"49","author":"Saulnier","year":"2021","journal-title":"Ann BioMed Eng."},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1002\/bdr2.1451","article-title":"Testicular function in cultured postnatal mouse testis fragments is similar to that of animals during the first wave of spermatogenesis","volume":"111","author":"Pence","year":"2019","journal-title":"Birth Defects Res"},{"key":"B40","doi-asserted-by":"publisher","first-page":"5380","DOI":"10.3390\/ijms20215380","article-title":"Vitamin e but not GSH decreases reactive oxygen species accumulation and enhances sperm production during In vitro maturation of frozen-thawed prepubertal mouse testicular tissue","volume":"20","author":"Arkoun","year":"2019","journal-title":"Int J Mol Sci"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2017.05.125","article-title":"The effects of IGF-1 on mouse spermatogenesis using an organ culture method","volume":"491","author":"Yao","year":"2017","journal-title":"Biochem Biophys Res Commun"},{"key":"B42","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1095\/biolreprod.113.114702","article-title":"Wt1 is involved in leydig cell steroid hormone biosynthesis by regulating paracrine factor expression in mice","volume":"90","author":"Chen","year":"2014","journal-title":"Biol Reprod"},{"key":"B43","doi-asserted-by":"publisher","first-page":"87","DOI":"10.1210\/mend.11.1.9871","article-title":"Repression of cAMP-induced expression of the mouse P450 17 alpha-hydroxylase\/C17-20 lyase gene (Cyp17) by androgens","volume":"11","author":"Burgos-Trinidad","year":"1997","journal-title":"Mol Endocrinol"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1262\/jrd.49.275","article-title":"Alterations of gene expression in adult male rat testis and pituitary shortly after subacute administration of the antiandrogen flutamide","volume":"49","author":"Ohsako","year":"2003","journal-title":"J Reprod Dev"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod.104.035915","article-title":"Androgen-regulated transcripts in the neonatal mouse testis as determined through microarray analysis","volume":"72","author":"Zhou","year":"2005","journal-title":"Biol Reprod"},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.115.17.3491","article-title":"Failure of normal adult leydig cell development in androgen-receptor-deficient mice","volume":"115","author":"O\u2019Shaughnessy","year":"2002","journal-title":"J Cell Sci"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1210\/en.2009-0577","article-title":"Opposing roles of insulin-like growth factor binding protein 3 and humanin in the regulation of testicular germ cell apoptosis","volume":"151","author":"Lue","year":"2010","journal-title":"Endocrinology."},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M109.046847","article-title":"Interaction of insulin-like growth factor-binding protein-3 and BAX in mitochondria promotes male germ cell apoptosis","volume":"285","author":"Jia","year":"2010","journal-title":"J Biol Chem"},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M002547200","article-title":"Direct functional interactions between insulin-like growth factor-binding protein-3 and retinoid X receptor-alpha regulate transcriptional signaling and apoptosis","volume":"275","author":"Liu","year":"2000","journal-title":"J Biol Chem"},{"key":"B50","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2003-07-2203","article-title":"Insulin-like growth factor binding protein-3 antagonizes the effects of retinoids in myeloid leukemia cells","volume":"104","author":"Ikezoe","year":"2004","journal-title":"Blood."},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1152\/ajpendo.90846.2008","article-title":"Inhibition of adipocyte differentiation by insulin-like growth factor-binding protein-3","volume":"296","author":"Chan","year":"2009","journal-title":"Am J Physiol Endocrinol Metab"},{"key":"B52","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1038\/onc.2012.538","article-title":"The role of insulin-like growth factor binding protein-3 in the breast cancer cell response to DNA-damaging agents","volume":"33","author":"Lin","year":"2014","journal-title":"Oncogene."},{"key":"B53","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2135111100","article-title":"Interaction between the Alzheimer's survival peptide humanin and insulin-like growth factor-binding protein 3 regulates cell survival and apoptosis","volume":"100","author":"Ikonen","year":"2003","journal-title":"Proc Natl Acad Sci USA"},{"key":"B54","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod.108.070839","article-title":"Levonorgestrel enhances spermatogenesis suppression by testosterone with greater alteration in testicular gene expression in men","volume":"80","author":"Lue","year":"2009","journal-title":"Biol Reprod"},{"key":"B55","doi-asserted-by":"publisher","DOI":"10.1093\/molehr\/gaw063","article-title":"Vitamin a prevents round spermatid nuclear damage and promotes the production of motile sperm during in vitro maturation of vitrified pre-pubertal mouse testicular tissue","volume":"22","author":"Dumont","year":"2016","journal-title":"Mol Hum Reprod"},{"key":"B56","doi-asserted-by":"publisher","DOI":"10.1023\/A:1016682520887","article-title":"Effective generation of very low density lipoprotein receptor transgenic mice by overlapping genomic DNA fragments: high testis expression and disturbed spermatogenesis","volume":"10","author":"Tacken","year":"2001","journal-title":"Transgenic Res"},{"key":"B57","doi-asserted-by":"publisher","DOI":"10.3177\/jnsv.35.655","article-title":"Changes in distribution of alpha-tocopherol and cholesterol in serum lipoproteins and tissues of rats by dietary PCB and dietary level of protein","volume":"35","author":"Kato","year":"1989","journal-title":"J Nutr Sci Vitaminol (Tokyo)."},{"key":"B58","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgen.1005863","article-title":"UPF2-dependent nonsense-mediated mRNA decay pathway is essential for spermatogenesis by selectively eliminating longer 3'UTR transcripts","volume":"12","author":"Bao","year":"2016","journal-title":"PLoS Genet"},{"key":"B59","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1186\/s13059-019-1633-3","article-title":"Gene editing of the multi-copy H2A.B gene and its importance for fertility","volume":"20","author":"Anuar","year":"2019","journal-title":"Genome Biol"},{"key":"B60","doi-asserted-by":"publisher","DOI":"10.1194\/jlr.R016675","article-title":"Early steps in steroidogenesis: intracellular cholesterol trafficking","volume":"52","author":"Miller","year":"2011","journal-title":"J Lipid Res"},{"key":"B61","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkv345","article-title":"The ReproGenomics viewer: an integrative cross-species toolbox for the reproductive science community","volume":"43","author":"Darde","year":"2015","journal-title":"Nucleic Acids Res"},{"key":"B62","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btz047","article-title":"The ReproGenomics viewer: a multi-omics and cross-species resource compatible with single-cell studies for the reproductive science community","volume":"35","author":"Darde","year":"2019","journal-title":"Bioinformatics."}],"container-title":["Frontiers in Endocrinology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fendo.2023.1112834\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,3,17]],"date-time":"2023-03-17T05:17:06Z","timestamp":1679030226000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fendo.2023.1112834\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,3,17]]},"references-count":62,"alternative-id":["10.3389\/fendo.2023.1112834"],"URL":"https:\/\/doi.org\/10.3389\/fendo.2023.1112834","relation":{},"ISSN":["1664-2392"],"issn-type":[{"value":"1664-2392","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,3,17]]},"article-number":"1112834"}}