{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,27]],"date-time":"2026-01-27T10:42:56Z","timestamp":1769510576600,"version":"3.49.0"},"reference-count":32,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2026,1,23]],"date-time":"2026-01-23T00:00:00Z","timestamp":1769126400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,1,26]],"date-time":"2026-01-26T00:00:00Z","timestamp":1769385600000},"content-version":"vor","delay-in-days":3,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100014584","name":"Universit\u00e4tsmedizin der Johannes Gutenberg-Universit\u00e4t Mainz","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100014584","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Fetuin-B is crucial in fertilization through inhibiting ovastacin, an enzyme that hardens the zona pellucida. Primarily synthesized in the liver, fetuin-B is also found in the placenta, tongue, and follicular fluid produced by granulosa cells. This study investigated fetuin-B localization and production in granulosa cells. Samples from 45 women undergoing intracytoplasmic sperm injection therapy at the University Medical Center Mainz were analyzed. Serum and follicular fluid samples were retrieved during egg collection. Granulosa cells were cultured and sampled at different time points.\n                    <jats:italic>FETUB<\/jats:italic>\n                    gene expression was quantified by RT-qPCR, with HepG2 cells as a positive control. Fetuin-B levels in granulosa cells and supernatants were measured by ELISA. Fetuin-B was localized using immunocytochemical staining. HepG2 cells showed strong\n                    <jats:italic>FETUB<\/jats:italic>\n                    expression, while granulosa cells exhibited minimal transcription. Fetuin-B levels were highest in serum and significantly lower in follicular fluid (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.001). Supernatant fetuin-B peaked on day 1 (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009=\u20090.004), while intracellular fetuin-B exceeded extracellular (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009=\u20090.008). Immunocytochemistry revealed a largely uniform intracellular distribution, with notable accumulations in filopodia. These findings indicate that granulosa cells store, release, and re-internalize fetuin-B but do not synthesize it. The early release at culture onset suggests a regulatory role in supporting fertilization conditions.\n                  <\/jats:p>","DOI":"10.1038\/s41598-026-36199-6","type":"journal-article","created":{"date-parts":[[2026,1,23]],"date-time":"2026-01-23T16:04:38Z","timestamp":1769184278000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Expression, transport, and storage of fetuin-B in human granulosa cells"],"prefix":"10.1038","volume":"16","author":[{"given":"Bartosz","family":"Linek","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ann-Christin","family":"Meyer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carmen","family":"Schoppe","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thorsten","family":"Enklaar","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dirk","family":"Prawitt","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Walburgis","family":"Brenner","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4099-1596","authenticated-orcid":false,"given":"Konstantin","family":"Hofmann","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2026,1,23]]},"reference":[{"key":"36199_CR1","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1042\/bj20030676","volume":"376","author":"B Denecke","year":"2003","unstructured":"Denecke, B. et al. Tissue distribution and activity testing suggest a similar but not identical function of fetuin-B and fetuin-A. Biochem. J. 376, 135\u2013145 (2003).","journal-title":"Biochem. J."},{"key":"36199_CR2","doi-asserted-by":"publisher","first-page":"963","DOI":"10.1172\/JCI41350","volume":"120","author":"JS Richards","year":"2010","unstructured":"Richards, J. S. & Pangas, S. A. The ovary: basic biology and clinical implications. J. Clin. Invest. 120, 963\u2013972 (2010).","journal-title":"J. Clin. Invest."},{"key":"36199_CR3","doi-asserted-by":"publisher","first-page":"2911","DOI":"10.2741\/3422","volume":"14","author":"C Lee","year":"2009","unstructured":"Lee, C., Bongcam-Rudloff, E., Sollner, C. & Jahnen-Dechent, W. Claesson-Welsh, L. Type 3 cystatins; fetuins, kininogen and histidine-rich glycoprotein. Front. Biosci. Landmark Ed. 14, 2911\u20132922 (2009).","journal-title":"Front. Biosci. Landmark Ed."},{"key":"36199_CR4","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1083\/jcb.201112094","volume":"197","author":"AD Burkart","year":"2012","unstructured":"Burkart, A. D., Xiong, B., Baibakov, B., Jim\u00e9nez-Movilla, M. & Dean, J. Ovastacin, a cortical granule protease, cleaves ZP2 in the Zona pellucida to prevent polyspermy. J. Cell. Biol. 197, 37\u201344 (2012).","journal-title":"J. Cell. Biol."},{"key":"36199_CR5","doi-asserted-by":"publisher","first-page":"1195","DOI":"10.1515\/hsz-2014-0189","volume":"395","author":"W St\u00f6cker","year":"2014","unstructured":"St\u00f6cker, W. et al. Mammalian gamete fusion depends on the Inhibition of ovastacin by fetuin-B. Biol. Chem. 395, 1195\u20131199 (2014).","journal-title":"Biol. Chem."},{"key":"36199_CR6","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1093\/molehr\/gaw067","volume":"23","author":"E Dietzel","year":"2017","unstructured":"Dietzel, E., Floehr, J., Van de Leur, E., Weiskirchen, R. & Jahnen-Dechent, W. Recombinant fetuin-B protein maintains high fertilization rate in cumulus cell-free mouse oocytes. Mol. Hum. Reprod. 23, 25\u201333 (2017).","journal-title":"Mol. Hum. Reprod."},{"key":"36199_CR7","first-page":"379","volume-title":"Current Topics in Developmental Biology","author":"SK Gupta","year":"2018","unstructured":"Gupta, S. K. Chapter Twelve - The Human Egg\u2019s Zona Pellucida. In Current Topics in Developmental Biology Vol. 130 (eds Litscher, E. S. & Wassarman, P. M.) 379\u2013411 (Academic, 2018)."},{"key":"36199_CR8","doi-asserted-by":"publisher","first-page":"326","DOI":"10.1002\/mrd.23320","volume":"87","author":"E Fahrenkamp","year":"2020","unstructured":"Fahrenkamp, E., Algarra, B. & Jovine, L. Mammalian egg coat modifications and the block to polyspermy. Mol. Reprod. Dev. 87, 326\u2013340 (2020).","journal-title":"Mol. Reprod. Dev."},{"key":"36199_CR9","doi-asserted-by":"publisher","first-page":"106","DOI":"10.1016\/j.devcel.2013.03.001","volume":"25","author":"E Dietzel","year":"2013","unstructured":"Dietzel, E. et al. Fetuin-B, a Liver-Derived plasma protein is essential for fertilization. Dev. Cell. 25, 106\u2013112 (2013).","journal-title":"Dev. Cell."},{"key":"36199_CR10","doi-asserted-by":"publisher","first-page":"630","DOI":"10.1093\/humrep\/dev340","volume":"31","author":"J Floehr","year":"2016","unstructured":"Floehr, J. et al. Association of high fetuin-B concentrations in serum with fertilization rate in IVF: a cross-sectional pilot study. Hum. Reprod. Oxf. Engl. 31, 630\u2013637 (2016).","journal-title":"Hum. Reprod. Oxf. Engl."},{"key":"36199_CR11","doi-asserted-by":"publisher","first-page":"819","DOI":"10.1387\/ijdb.120133cc","volume":"56","author":"M Hennet","year":"2013","unstructured":"Hennet, M. & Combelles, C. M. H. The antral follicle: a microenvironment for oocyte differentiation. Int. J. Dev. Biol. 56, 819\u2013831 (2013).","journal-title":"Int. J. Dev. Biol."},{"key":"36199_CR12","doi-asserted-by":"publisher","first-page":"543","DOI":"10.1095\/biolreprod43.4.543","volume":"43","author":"R Buccione","year":"1990","unstructured":"Buccione, R., Schroeder, A. C. & Eppig, J. J. Interactions between somatic cells and germ cells throughout mammalian oogenesis. Biol. Reprod. 43, 543\u2013547 (1990).","journal-title":"Biol. Reprod."},{"key":"36199_CR13","doi-asserted-by":"publisher","first-page":"1418","DOI":"10.3390\/cells9061418","volume":"9","author":"M Jozkowiak","year":"2020","unstructured":"Jozkowiak, M. et al. The stemness of human ovarian granulosa cells and the role of Resveratrol in the differentiation of MSCs-A review based on cellular and molecular knowledge. Cells 9, 1418 (2020).","journal-title":"Cells"},{"key":"36199_CR14","volume-title":"Endotext","author":"CJ Williams","year":"2000","unstructured":"Williams, C. J. & Erickson, G. F. Morphology and physiology of the ovary. In Endotext (eds Feingold, K. R. et al.) (MDText.com, Inc, 2000)."},{"key":"36199_CR15","doi-asserted-by":"publisher","first-page":"P215","DOI":"10.1055\/s-0036-1592764","volume":"76","author":"B Linek","year":"2016","unstructured":"Linek, B. et al. Fetuin-B \u2013 ein Neuer marker f\u00fcr eine gelungene IVF? Geburtshilfe Frauenheilkd. 76, P215 (2016).","journal-title":"Geburtshilfe Frauenheilkd"},{"key":"36199_CR16","first-page":"1498","volume":"11","author":"W Zhou","year":"2019","unstructured":"Zhou, W. et al. Fetuin B aggravates liver X receptor-mediated hepatic steatosis through AMPK in HepG2 cells and mice. Am. J. Transl Res. 11, 1498\u20131509 (2019).","journal-title":"Am. J. Transl Res."},{"key":"36199_CR17","unstructured":"FETUB fetuin B. [Homo sapiens (human)] - Gene - NCBI.\u00a0https:\/\/www-ncbi-nlm-nih-gov.proxy.ub.uni-frankfurt.de\/gene\/26998"},{"key":"36199_CR18","unstructured":"WMA - The World Medical Association-WMA Declaration of Helsinki \u2013 Ethical Principles for Medical Research. Involving Human Subjects.\u00a0https:\/\/www.wma.net\/policies-post\/wma-declaration-of-helsinki-ethical-principles-for-medical-research-involving-human-subjects\/"},{"key":"36199_CR19","doi-asserted-by":"publisher","first-page":"S531","DOI":"10.33549\/physiolres.934017","volume":"67","author":"P \u0160imj\u00e1k","year":"2018","unstructured":"\u0160imj\u00e1k, P. et al. Changes in plasma concentrations and mRNA expression of hepatokines fetuin A, fetuin B and FGF21 in physiological pregnancy and gestational diabetes mellitus. Physiol. Res. 67, S531\u2013S542 (2018).","journal-title":"Physiol. Res."},{"key":"36199_CR20","doi-asserted-by":"publisher","first-page":"1655","DOI":"10.1095\/biolreprod65.6.1655","volume":"65","author":"PE H\u00f8yer","year":"2001","unstructured":"H\u00f8yer, P. E., Terkelsen, O. B. F., Byskov, G., Nielsen, H. & A. & Fetuin and fetuin messenger RNA in granulosa cells of the rat Ovary1. Biol. Reprod. 65, 1655\u20131662 (2001).","journal-title":"Biol. Reprod."},{"key":"36199_CR21","doi-asserted-by":"publisher","first-page":"149","DOI":"10.1016\/0167-4781(92)90522-2","volume":"1130","author":"F Yang","year":"1992","unstructured":"Yang, F., Chen, Z. L., Bergeron, J. M., Cupples, R. L. & Friedrichs, W. E. Human alpha 2-HS-glycoprotein\/bovine fetuin homologue in mice: identification and developmental regulation of the gene. Biochim. Biophys. Acta. 1130, 149\u2013156 (1992).","journal-title":"Biochim. Biophys. Acta"},{"key":"36199_CR22","doi-asserted-by":"publisher","first-page":"719029","DOI":"10.3389\/fendo.2021.719029","volume":"12","author":"R Punchoo","year":"2021","unstructured":"Punchoo, R. & Bhoora, S. Variation in the measurement of Anti-M\u00fcllerian Hormone \u2013 What are the laboratory issues? Front. Endocrinol. 12, 719029 (2021).","journal-title":"Front. Endocrinol."},{"key":"36199_CR23","doi-asserted-by":"publisher","first-page":"208","DOI":"10.1093\/molehr\/gat078","volume":"20","author":"M B\u00e4chler","year":"2014","unstructured":"B\u00e4chler, M., Menshykau, D., De Geyter, C. & Iber, D. Species-specific differences in follicular antral sizes result from diffusion-based limitations on the thickness of the granulosa cell layer. Mol. Hum. Reprod. 20, 208\u2013221 (2014).","journal-title":"Mol. Hum. Reprod."},{"key":"36199_CR24","unstructured":"E, D. & W, J. D. J, F. The biological role of Fetuin-B in female reproduction. Ann Reprod. Med. Treat\u00a0https:\/\/www.jscimedcentral.com\/article\/The-Biological-Role-of-Fetuin-B--in-Female-Reproduction\u00a0(2016)."},{"key":"36199_CR25","first-page":"285","volume":"46","author":"X Hu","year":"2017","unstructured":"Hu, X. et al. [Correlation of fetuin-B concentrations in serum and follicular fluid with outcomes of in vitro fertilization]. Zhejiang Xue Xue Bao Yi Xue Ban J. Zhejiang Univ. Med. Sci. 46, 285\u2013289 (2017).","journal-title":"Zhejiang Xue Xue Bao Yi Xue Ban J. Zhejiang Univ. Med. Sci."},{"key":"36199_CR26","doi-asserted-by":"publisher","first-page":"1313","DOI":"10.1515\/cclm-2014-0211","volume":"52","author":"J B\u00f3dis","year":"2014","unstructured":"B\u00f3dis, J., Peti, A. M., Sulyok, E., Kov\u00e1cs, G. L. & V\u00e1rnagy, A. Serum and follicular fluid fetuin-A in women undergoing in vitro fertilization. Clin. Chem. Lab. Med. 52, 1313\u20131318 (2014).","journal-title":"Clin. Chem. Lab. Med."},{"key":"36199_CR27","doi-asserted-by":"publisher","first-page":"798","DOI":"10.1093\/oxfordjournals.humrep.a019257","volume":"11","author":"D Spitzer","year":"1996","unstructured":"Spitzer, D., Murach, K. F., Lottspeich, F., Staudach, A. & Illmensee, K. Different protein patterns derived from follicular fluid of mature and immature human follicles. Hum. Reprod. Oxf. Engl. 11, 798\u2013807 (1996).","journal-title":"Hum. Reprod. Oxf. Engl."},{"key":"36199_CR28","doi-asserted-by":"publisher","first-page":"1101","DOI":"10.1007\/s10815-019-01454-5","volume":"36","author":"L Fang","year":"2019","unstructured":"Fang, L. et al. Serum and follicular fluid fetuin-B levels are correlated with fertilization rates in conventional IVF cycles. J. Assist. Reprod. Genet. 36, 1101\u20131107 (2019).","journal-title":"J. Assist. Reprod. Genet."},{"key":"36199_CR29","doi-asserted-by":"publisher","first-page":"1989","DOI":"10.3390\/life13101989","volume":"13","author":"R Sciorio","year":"1989","unstructured":"Sciorio, R., Cariati, F., Fleming, S. & Alviggi, C. Exploring the Impact of Controlled Ovarian Stimulation and Non-Invasive Oocyte Assessment in ART Treatments. Life 13, 1989 (1989).","journal-title":"Life"},{"key":"36199_CR30","doi-asserted-by":"publisher","first-page":"S19","DOI":"10.1016\/j.ejogrb.2004.01.010","volume":"115","author":"S Cecconi","year":"2004","unstructured":"Cecconi, S., Ciccarelli, C., Barberi, M., Macchiarelli, G. & Canipari, R. Granulosa cell-oocyte interactions. Eur. J. Obstet. Gynecol. Reprod. Biol. 115, S19\u2013S22 (2004).","journal-title":"Eur. J. Obstet. Gynecol. Reprod. Biol."},{"key":"36199_CR31","doi-asserted-by":"publisher","first-page":"217","DOI":"10.1055\/s-2006-948551","volume":"24","author":"DL Russell","year":"2006","unstructured":"Russell, D. L. & Salustri, A. Extracellular matrix of the cumulus-oocyte complex. Semin Reprod. Med. 24, 217\u2013227 (2006).","journal-title":"Semin Reprod. Med."},{"key":"36199_CR32","doi-asserted-by":"publisher","first-page":"q1591","DOI":"10.1136\/bmj.q1591","volume":"386","author":"Z Kmietowicz","year":"2024","unstructured":"Kmietowicz, Z. Average age of starting fertility treatment rises to over 35 in UK. BMJ 386, q1591 (2024).","journal-title":"BMJ"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-026-36199-6","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-026-36199-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-026-36199-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,26]],"date-time":"2026-01-26T23:02:32Z","timestamp":1769468552000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-026-36199-6"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,23]]},"references-count":32,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,12]]}},"alternative-id":["36199"],"URL":"https:\/\/doi.org\/10.1038\/s41598-026-36199-6","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,23]]},"assertion":[{"value":"28 January 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"9 January 2026","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"23 January 2026","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}},{"value":"During the preparation of this work the authors used ChatGPT 3.5 (OpenAI; USA) to increase readability. After using this tool\/service, the authors reviewed and edited the content as needed and take full responsibility for the content of the published article.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declaration of generative AI and AI-assisted technologies in the writing process"}}],"article-number":"3264"}}