{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,21]],"date-time":"2026-04-21T07:36:58Z","timestamp":1776757018926,"version":"3.51.2"},"reference-count":41,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2026,1,20]],"date-time":"2026-01-20T00:00:00Z","timestamp":1768867200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,1,20]],"date-time":"2026-01-20T00:00:00Z","timestamp":1768867200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Cell Regen"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The placenta plays a pivotal role in human pregnancy, yet research into placental development has been hindered by limited access to early-stage embryos and ethical constraints. Although human trophoblast stem cells (hTSCs) have been established from blastocysts, deriving these cells efficiently from primed human pluripotent stem cells (hPSCs) remains challenging. Here, we developed a simplified and efficient strategy that enables direct, efficient conversion of primed hPSCs into stable, self-renewing hTSCs by transiently inhibiting the MEK\/ERK signaling pathway using the inhibitor PD0325901 in a simplified basal medium. This approach significantly enhanced the generation of trophoblast cells expressing the critical trophoblast marker GATA3 and led to the establishment of homogeneous hTSC lines with robust capacities to differentiate into functional extravillous trophoblast (EVT) and syncytiotrophoblast (STB) lineages. Transcriptomic and chromatin accessibility analyses confirmed that these hTSCs closely resembled blastocyst-derived trophoblast cells and clearly differed from amnion lineages, confirming authentic trophoblast identity distinct from amnion. Additionally, precise modulation of WNT signaling activity was essential for optimal trophoblast induction efficiency, highlighting the importance of signaling equilibrium in trophoblast differentiation. Collectively, our optimized protocol offers an accessible and reproducible platform for modeling early placental development and understanding the pathogenesis of trophoblast-associated disorders in vitro.<\/jats:p>","DOI":"10.1186\/s13619-025-00261-x","type":"journal-article","created":{"date-parts":[[2026,1,20]],"date-time":"2026-01-20T02:01:01Z","timestamp":1768874461000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Transient inhibition of MEK\/ERK and WNT pathways enhances direct differentiation of primed hPSCs into functional trophoblast stem cells"],"prefix":"10.1186","volume":"15","author":[{"given":"Qifan","family":"Jiang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ping","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chunlin","family":"Chen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2026,1,20]]},"reference":[{"key":"261_CR1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.202778","author":"PA Balestrini","year":"2024","unstructured":"Balestrini PA, Abdelbaki A, McCarthy A, Devito L, Senner CE, Chen AE, et al. Transcription factor-based transdifferentiation of human embryonic to trophoblast stem cells. Development. 2024. https:\/\/doi.org\/10.1242\/dev.202778.","journal-title":"Development"},{"key":"261_CR2","doi-asserted-by":"publisher","first-page":"134","DOI":"10.1016\/j.placenta.2017.06.007","volume":"60","author":"GJ Burton","year":"2017","unstructured":"Burton GJ, Jauniaux E. The cytotrophoblastic shell and complications of pregnancy. Placenta. 2017;60:134\u20139. https:\/\/doi.org\/10.1016\/j.placenta.2017.06.007.","journal-title":"Placenta"},{"issue":"3","key":"261_CR3","doi-asserted-by":"publisher","first-page":"C664","DOI":"10.1152\/ajpcell.00295.2019","volume":"318","author":"Z Chen","year":"2020","unstructured":"Chen Z, Zhang J, Yuan A, Han J, Tan L, Zhou Z, et al. R-spondin3 promotes the tumor growth of choriocarcinoma JEG-3 cells. Am J Physiol Cell Physiol. 2020;318(3):C664\u201374. https:\/\/doi.org\/10.1152\/ajpcell.00295.2019.","journal-title":"Am J Physiol Cell Physiol"},{"key":"261_CR4","doi-asserted-by":"publisher","unstructured":"Dattani A, Corujo-Simon E, Radley A, Heydari T, Taheriabkenar Y, Carlisle F, et al. Naive pluripotent stem cell-based models capture FGF-dependent human hypoblast lineage specification. Cell Stem Cell. 2024;31(7):1058\u201371.e5. https:\/\/doi.org\/10.1016\/j.stem.2024.05.003.","DOI":"10.1016\/j.stem.2024.05.003"},{"key":"261_CR5","doi-asserted-by":"publisher","DOI":"10.1242\/dev.172940","author":"J Deathridge","year":"2019","unstructured":"Deathridge J, Antolovi\u0107 V, Parsons M, Chubb JR. Live imaging of ERK signalling dynamics in differentiating mouse embryonic stem cells. Development. 2019. https:\/\/doi.org\/10.1242\/dev.172940.","journal-title":"Development"},{"key":"261_CR6","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.52504","author":"C Dong","year":"2020","unstructured":"Dong C, Beltcheva M, Gontarz P, Zhang B, Popli P, Fischer LA, et al. Derivation of trophoblast stem cells from na\u00efve human pluripotent stem cells. Elife. 2020. https:\/\/doi.org\/10.7554\/eLife.52504.","journal-title":"Elife"},{"key":"261_CR7","doi-asserted-by":"publisher","first-page":"91","DOI":"10.1007\/978-1-0716-1908-7_7","volume":"2416","author":"C Dong","year":"2022","unstructured":"Dong C, Theunissen TW. Generating Trophoblast Stem Cells from Human Na\u00efve Pluripotent Stem Cells. Methods in Molecular Biology (Clifton, NJ). 2022;2416:91\u2013104. https:\/\/doi.org\/10.1007\/978-1-0716-1908-7_7.","journal-title":"Methods in Molecular Biology (Clifton, NJ)"},{"issue":"5","key":"261_CR8","doi-asserted-by":"publisher","first-page":"443","DOI":"10.1111\/j.1432-0436.2007.00249.x","volume":"76","author":"NS Funa","year":"2008","unstructured":"Funa NS, Saldeen J, Akerblom B, Welsh M. Interdependent fibroblast growth factor and activin A signaling promotes the expression of endodermal genes in differentiating mouse embryonic stem cells expressing Src homology 2-domain inactive Shb. Differentiation. 2008;76(5):443\u201353. https:\/\/doi.org\/10.1111\/j.1432-0436.2007.00249.x.","journal-title":"Differentiation"},{"issue":"7479","key":"261_CR9","doi-asserted-by":"publisher","first-page":"282","DOI":"10.1038\/nature12745","volume":"504","author":"O Gafni","year":"2013","unstructured":"Gafni O, Weinberger L, Mansour AA, Manor YS, Chomsky E, Ben-Yosef D, et al. Derivation of novel human ground state naive pluripotent stem cells. Nature. 2013;504(7479):282\u20136. https:\/\/doi.org\/10.1038\/nature12745.","journal-title":"Nature"},{"issue":"6","key":"261_CR10","doi-asserted-by":"publisher","first-page":"1040","DOI":"10.1016\/j.stem.2021.02.025","volume":"28","author":"G Guo","year":"2021","unstructured":"Guo G, Stirparo GG, Strawbridge SE, Spindlow D, Yang J, Clarke J, et al. Human naive epiblast cells possess unrestricted lineage potential. Cell Stem Cell. 2021;28(6):1040-56.e6. https:\/\/doi.org\/10.1016\/j.stem.2021.02.025.","journal-title":"Cell Stem Cell"},{"issue":"13","key":"261_CR11","doi-asserted-by":"publisher","first-page":"1501","DOI":"10.1089\/scd.2013.0329","volume":"23","author":"X Huang","year":"2014","unstructured":"Huang X, Han X, Uyunbilig B, Zhang M, Duo S, Zuo Y, et al. Establishment of bovine trophoblast stem-like cells from in vitro-produced blastocyst-stage embryos using two inhibitors. Stem Cells Dev. 2014;23(13):1501\u201314. https:\/\/doi.org\/10.1089\/scd.2013.0329.","journal-title":"Stem Cells Dev"},{"issue":"6","key":"261_CR12","doi-asserted-by":"publisher","first-page":"1023","DOI":"10.1016\/j.stem.2021.03.013","volume":"28","author":"S Io","year":"2021","unstructured":"Io S, Kabata M, Iemura Y, Semi K, Morone N, Minagawa A, et al. Capturing human trophoblast development with naive pluripotent stem cells in vitro. Cell Stem Cell. 2021;28(6):1023-39.e13. https:\/\/doi.org\/10.1016\/j.stem.2021.03.013.","journal-title":"Cell Stem Cell"},{"issue":"5","key":"261_CR13","doi-asserted-by":"publisher","first-page":"327","DOI":"10.1016\/j.placenta.2012.01.020","volume":"33","author":"JL James","year":"2012","unstructured":"James JL, Carter AM, Chamley LW. Human placentation from nidation to 5 weeks of gestation. Part I: what do we know about formative placental development following implantation? Placenta. 2012;33(5):327\u201334. https:\/\/doi.org\/10.1016\/j.placenta.2012.01.020.","journal-title":"Placenta"},{"issue":"20","key":"261_CR14","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2115709119","volume":"119","author":"YJ Jang","year":"2022","unstructured":"Jang YJ, Kim M, Lee BK, Kim J. Induction of human trophoblast stem-like cells from primed pluripotent stem cells. Proc Natl Acad Sci U S A. 2022;119(20):e2115709119. https:\/\/doi.org\/10.1073\/pnas.2115709119.","journal-title":"Proc Natl Acad Sci U S A"},{"key":"261_CR15","doi-asserted-by":"publisher","unstructured":"Karvas RM, Khan SA, Verma S, Yin Y, Kulkarni D, Dong C, et al. Stem-cell-derived trophoblast organoids model human placental development and susceptibility to emerging pathogens. Cell Stem Cell. 2022;29(5):810\u201325.e8. https:\/\/doi.org\/10.1016\/j.stem.2022.04.004.","DOI":"10.1016\/j.stem.2022.04.004"},{"issue":"10","key":"261_CR16","doi-asserted-by":"publisher","first-page":"1307","DOI":"10.1002\/stem.3052","volume":"37","author":"CQE Lee","year":"2019","unstructured":"Lee CQE, Bailey A, Lopez-Tello J, Sferruzzi-Perri AN, Okkenhaug K, Moffett A, et al. Inhibition of phosphoinositide-3-kinase signaling promotes the stem cell state of trophoblast. Stem Cells. 2019;37(10):1307\u201318. https:\/\/doi.org\/10.1002\/stem.3052.","journal-title":"Stem Cells"},{"issue":"Suppl","key":"261_CR17","doi-asserted-by":"publisher","first-page":"S28","DOI":"10.1016\/j.placenta.2011.12.003","volume":"33","author":"RM Lewis","year":"2012","unstructured":"Lewis RM, Cleal JK, Hanson MA. Review: Placenta, evolution and lifelong health. Placenta. 2012;33(Suppl):S28-32. https:\/\/doi.org\/10.1016\/j.placenta.2011.12.003.","journal-title":"Placenta"},{"issue":"10","key":"261_CR18","doi-asserted-by":"publisher","first-page":"1481","DOI":"10.1101\/gr.233437.117","volume":"28","author":"D Liu","year":"2018","unstructured":"Liu D, Wang X, He D, Sun C, He X, Yan L, et al. Single-cell RNA-sequencing reveals the existence of naive and primed pluripotency in pre-implantation rhesus monkey embryos. Genome Res. 2018;28(10):1481\u201393. https:\/\/doi.org\/10.1101\/gr.233437.117.","journal-title":"Genome Res"},{"issue":"11","key":"261_CR19","doi-asserted-by":"publisher","first-page":"1055","DOI":"10.1038\/nmeth.4436","volume":"14","author":"X Liu","year":"2017","unstructured":"Liu X, Nefzger CM, Rossello FJ, Chen J, Knaupp AS, Firas J, et al. Comprehensive characterization of distinct states of human naive pluripotency generated by reprogramming. Nat Methods. 2017;14(11):1055\u201362. https:\/\/doi.org\/10.1038\/nmeth.4436.","journal-title":"Nat Methods"},{"issue":"3","key":"261_CR20","doi-asserted-by":"publisher","first-page":"151","DOI":"10.1016\/j.exphem.2015.12.008","volume":"44","author":"X Ma","year":"2016","unstructured":"Ma X, Chen H, Chen L. A dual role of Erk signaling in embryonic stem cells. Exp Hematol. 2016;44(3):151\u20136. https:\/\/doi.org\/10.1016\/j.exphem.2015.12.008.","journal-title":"Exp Hematol"},{"key":"261_CR21","doi-asserted-by":"publisher","DOI":"10.1093\/molehr\/gaac032","author":"T Mizutani","year":"2022","unstructured":"Mizutani T, Orisaka M, Miyazaki Y, Morichika R, Uesaka M, Miyamoto K, et al. Inhibition of YAP\/TAZ-TEAD activity induces cytotrophoblast differentiation into syncytiotrophoblast in human trophoblast. Mol Hum Reprod. 2022. https:\/\/doi.org\/10.1093\/molehr\/gaac032.","journal-title":"Mol Hum Reprod"},{"key":"261_CR22","doi-asserted-by":"publisher","first-page":"79","DOI":"10.3389\/fcell.2020.00079","volume":"8","author":"M Mossahebi-Mohammadi","year":"2020","unstructured":"Mossahebi-Mohammadi M, Quan M, Zhang JS, Li X. FGF signaling pathway: a key regulator of stem cell pluripotency. Frontiers in Cell and Developmental Biology. 2020;8:79. https:\/\/doi.org\/10.3389\/fcell.2020.00079.","journal-title":"Frontiers in Cell and Developmental Biology"},{"issue":"6","key":"261_CR23","doi-asserted-by":"publisher","first-page":"985","DOI":"10.1016\/j.cell.2017.05.016","volume":"169","author":"R Nusse","year":"2017","unstructured":"Nusse R, Clevers H. Wnt\/\u03b2-catenin signaling, disease, and emerging therapeutic modalities. Cell. 2017;169(6):985\u201399. https:\/\/doi.org\/10.1016\/j.cell.2017.05.016.","journal-title":"Cell"},{"issue":"1","key":"261_CR24","doi-asserted-by":"publisher","first-page":"50","DOI":"10.1016\/j.stem.2017.11.004","volume":"22","author":"H Okae","year":"2018","unstructured":"Okae H, Toh H, Sato T, Hiura H, Takahashi S, Shirane K, et al. Derivation of Human Trophoblast Stem Cells. Cell Stem Cell. 2018;22(1):50-63.e6. https:\/\/doi.org\/10.1016\/j.stem.2017.11.004.","journal-title":"Cell Stem Cell"},{"issue":"7983","key":"261_CR25","doi-asserted-by":"publisher","first-page":"562","DOI":"10.1038\/s41586-023-06604-5","volume":"622","author":"B Oldak","year":"2023","unstructured":"Oldak B, Wildschutz E, Bondarenko V, Comar MY, Zhao C, Aguilera-Castrejon A, et al. Complete human day 14 post-implantation embryo models from naive ES cells. Nature. 2023;622(7983):562\u201373. https:\/\/doi.org\/10.1038\/s41586-023-06604-5.","journal-title":"Nature"},{"issue":"3","key":"261_CR26","doi-asserted-by":"publisher","first-page":"393","DOI":"10.1016\/j.mcn.2008.03.012","volume":"38","author":"SM Pollard","year":"2008","unstructured":"Pollard SM, Wallbank R, Tomlinson S, Grotewold L, Smith A. Fibroblast growth factor induces a neural stem cell phenotype in foetal forebrain progenitors and during embryonic stem cell differentiation. Mol Cell Neurosci. 2008;38(3):393\u2013403. https:\/\/doi.org\/10.1016\/j.mcn.2008.03.012.","journal-title":"Mol Cell Neurosci"},{"issue":"3","key":"261_CR27","doi-asserted-by":"publisher","first-page":"313","DOI":"10.1016\/j.bpobgyn.2011.02.006","volume":"25","author":"R Romero","year":"2011","unstructured":"Romero R, Kusanovic JP, Chaiworapongsa T, Hassan SS. Placental bed disorders in preterm labor, preterm PROM, spontaneous abortion and abruptio placentae. Best Pract Res Clin Obstet Gynaecol. 2011;25(3):313\u201327. https:\/\/doi.org\/10.1016\/j.bpobgyn.2011.02.006.","journal-title":"Best Pract Res Clin Obstet Gynaecol"},{"issue":"4","key":"261_CR28","doi-asserted-by":"publisher","first-page":"874","DOI":"10.1634\/stemcells.2007-0608","volume":"26","author":"N Shiraki","year":"2008","unstructured":"Shiraki N, Yoshida T, Araki K, Umezawa A, Higuchi Y, Goto H, et al. Guided differentiation of embryonic stem cells into Pdx1-expressing regional-specific definitive endoderm. Stem Cells. 2008;26(4):874\u201385. https:\/\/doi.org\/10.1634\/stemcells.2007-0608.","journal-title":"Stem Cells"},{"issue":"40","key":"261_CR29","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2403003121","volume":"121","author":"V Shukla","year":"2024","unstructured":"Shukla V, Moreno-Irusta A, Varberg KM, Kuna M, Iqbal K, Galligos AM, et al. Notum-mediated WNT silencing drives extravillous trophoblast cell lineage development. Proc Natl Acad Sci U S A. 2024;121(40):e2403003121. https:\/\/doi.org\/10.1073\/pnas.2403003121.","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"12","key":"261_CR30","doi-asserted-by":"publisher","DOI":"10.1002\/bies.202400108","volume":"46","author":"A Smith","year":"2024","unstructured":"Smith A. Propagating pluripotency - the conundrum of self-renewal. BioEssays. 2024;46(12):e2400108. https:\/\/doi.org\/10.1002\/bies.202400108.","journal-title":"BioEssays"},{"issue":"7511","key":"261_CR31","doi-asserted-by":"publisher","first-page":"611","DOI":"10.1038\/nature13581","volume":"511","author":"ZD Smith","year":"2014","unstructured":"Smith ZD, Chan MM, Humm KC, Karnik R, Mekhoubad S, Regev A, et al. DNA methylation dynamics of the human preimplantation embryo. Nature. 2014;511(7511):611\u20135. https:\/\/doi.org\/10.1038\/nature13581.","journal-title":"Nature"},{"issue":"6","key":"261_CR32","doi-asserted-by":"publisher","first-page":"1303","DOI":"10.1016\/j.stemcr.2022.04.013","volume":"17","author":"F Soncin","year":"2022","unstructured":"Soncin F, Morey R, Bui T, Requena DF, Cheung VC, Kallol S, et al. Derivation of functional trophoblast stem cells from primed human pluripotent stem cells. Stem Cell Reports. 2022;17(6):1303\u201317. https:\/\/doi.org\/10.1016\/j.stemcr.2022.04.013.","journal-title":"Stem Cell Reports"},{"key":"261_CR33","doi-asserted-by":"publisher","DOI":"10.1242\/dev.163428","author":"MY Turco","year":"2019","unstructured":"Turco MY, Moffett A. Development of the human placenta. Development. 2019. https:\/\/doi.org\/10.1242\/dev.163428.","journal-title":"Development"},{"issue":"11","key":"261_CR34","doi-asserted-by":"publisher","first-page":"2484","DOI":"10.1016\/j.stemcr.2022.09.008","volume":"17","author":"S Viukov","year":"2022","unstructured":"Viukov S, Shani T, Bayerl J, Aguilera-Castrejon A, Oldak B, Sheban D, et al. Human primed and na\u00efve PSCs are both able to differentiate into trophoblast stem cells. Stem Cell Reports. 2022;17(11):2484\u2013500. https:\/\/doi.org\/10.1016\/j.stemcr.2022.09.008.","journal-title":"Stem Cell Reports"},{"issue":"1","key":"261_CR35","doi-asserted-by":"publisher","first-page":"1626","DOI":"10.1038\/s41467-022-29312-6","volume":"13","author":"LJ Wang","year":"2022","unstructured":"Wang LJ, Chen CP, Lee YS, Ng PS, Chang GD, Pao YH, et al. Functional antagonism between \u0394Np63\u03b1 and GCM1 regulates human trophoblast stemness and differentiation. Nat Commun. 2022;13(1):1626. https:\/\/doi.org\/10.1038\/s41467-022-29312-6.","journal-title":"Nat Commun"},{"key":"261_CR36","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abf4416","author":"Y Wei","year":"2021","unstructured":"Wei Y, Wang T, Ma L, Zhang Y, Zhao Y, Lye K, et al. Efficient derivation of human trophoblast stem cells from primed pluripotent stem cells. Sci Adv. 2021. https:\/\/doi.org\/10.1126\/sciadv.abf4416.","journal-title":"Sci Adv"},{"issue":"7194","key":"261_CR37","doi-asserted-by":"publisher","first-page":"524","DOI":"10.1038\/nature06894","volume":"453","author":"L Yang","year":"2008","unstructured":"Yang L, Soonpaa MH, Adler ED, Roepke TK, Kattman SJ, Kennedy M, et al. Human cardiovascular progenitor cells develop from a KDR+ embryonic-stem-cell-derived population. Nature. 2008;453(7194):524\u20138. https:\/\/doi.org\/10.1038\/nature06894.","journal-title":"Nature"},{"issue":"3","key":"261_CR38","doi-asserted-by":"publisher","first-page":"317","DOI":"10.1016\/j.devcel.2006.01.002","volume":"10","author":"W Yang","year":"2006","unstructured":"Yang W, Klaman LD, Chen B, Araki T, Harada H, Thomas SM, et al. An Shp2\/SFK\/Ras\/Erk signaling pathway controls trophoblast stem cell survival. Dev Cell. 2006;10(3):317\u201327. https:\/\/doi.org\/10.1016\/j.devcel.2006.01.002.","journal-title":"Dev Cell"},{"issue":"1","key":"261_CR39","doi-asserted-by":"publisher","first-page":"2","DOI":"10.1186\/s13287-017-0750-8","volume":"9","author":"Y Yu","year":"2018","unstructured":"Yu Y, Wang X, Zhang X, Zhai Y, Lu X, Ma H, et al. Erk inhibition promotes neuroectodermal precursor commitment by blocking self-renewal and primitive streak formation of the epiblast. Stem Cell Res Ther. 2018;9(1):2. https:\/\/doi.org\/10.1186\/s13287-017-0750-8.","journal-title":"Stem Cell Res Ther"},{"issue":"8","key":"261_CR40","doi-asserted-by":"publisher","first-page":"1219","DOI":"10.1016\/j.scib.2025.01.063","volume":"70","author":"W Zhang","year":"2025","unstructured":"Zhang W, Wang Q, Sun S, Liu Y, Zhao Y, Luo H, et al. Derivation and genetic-screening of human haploid trophoblast stem cells. Sci Bull. 2025;70(8):1219\u201323. https:\/\/doi.org\/10.1016\/j.scib.2025.01.063.","journal-title":"Sci Bull"},{"issue":"4","key":"261_CR41","doi-asserted-by":"publisher","DOI":"10.15252\/embr.202255235","volume":"24","author":"I Zorzan","year":"2023","unstructured":"Zorzan I, Betto RM, Rossignoli G, Arboit M, Drusin A, Corridori C, et al. Chemical conversion of human conventional PSCs to TSCs following transient naive gene activation. EMBO Rep. 2023;24(4):e55235. https:\/\/doi.org\/10.15252\/embr.202255235.","journal-title":"EMBO Rep"}],"container-title":["Cell Regeneration"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13619-025-00261-x.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s13619-025-00261-x","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13619-025-00261-x.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,20]],"date-time":"2026-01-20T02:01:02Z","timestamp":1768874462000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1186\/s13619-025-00261-x"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,20]]},"references-count":41,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,12]]}},"alternative-id":["261"],"URL":"https:\/\/doi.org\/10.1186\/s13619-025-00261-x","relation":{},"ISSN":["2045-9769"],"issn-type":[{"value":"2045-9769","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,20]]},"assertion":[{"value":"13 May 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"4 September 2025","order":2,"name":"revised","label":"Revised","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"8 September 2025","order":3,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"20 January 2026","order":4,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Not applicable.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare that they have no conflicts of interest.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"4"}}