{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T03:14:14Z","timestamp":1788146054130,"version":"build-2803163510"},"reference-count":46,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2026,8,30]],"date-time":"2026-08-30T00:00:00Z","timestamp":1788048000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Key Science-Technology Project of Inner Mongolia","award":["2023KYPT0010"],"award-info":[{"award-number":["2023KYPT0010"]}]},{"name":"Key Science-Technology Project of Inner Mongolia","award":["2026CXPT02003"],"award-info":[{"award-number":["2026CXPT02003"]}]},{"name":"STI 2030-Major Projects of China","award":["2023ZD0407504"],"award-info":[{"award-number":["2023ZD0407504"]}]},{"name":"Development Plan for Young Scientific and Technological Talents in Colleges and Universities of Inner Mongolia Autonomous Region of China","award":["NMGIRT2204"],"award-info":[{"award-number":["NMGIRT2204"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32160172"],"award-info":[{"award-number":["32160172"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["32160172"],"award-info":[{"award-number":["32160172"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Methyl-CpG binding domain protein 3 (MBD3) is a member of the nucleosome remodeling and deacetylase (NuRD) corepressor protein complex, which plays a pivotal role in embryonic development, pluripotent stem cell (PSC) differentiation, and induced pluripotent stem cell (iPSC) reprogramming. However, whether MBD3 is also involved in iPSC differentiation into primordial germ cell-like cells (PGCLCs) is unclear. In this study, bovine iPSCs generated by MBD3 knockdown (MBD3 KD) during reprogramming were subjected to PGCLC differentiation using a monolayer cell culture method. Our results revealed that MBD3 KD enhanced the ability of bovine iPSCs to differentiate into PGCLCs. MBD3-KD bovine iPSC-derived PGCLCs exhibited the characteristics of in vivo primordial germ cells (PGCs) in a migratory state. MBD3 KD increased NODAL signal transduction during the induction of PGCLCs and was associated with increased expression of PRDM1 and SOX15. Moreover, MBD3 KD increased the bivalent modification of transforming growth factor beta (TGF-\u03b2)-related genes concomitant with the upregulation of DPPA4 and DNMT3L expression in bovine iPSCs, and this epigenetic change was associated with enhanced NODAL signaling. Our results indicate that MBD3 KD during reprogramming favors the differentiation of bovine iPSCs into PGCLCs, further highlighting the impact of MBD3 on histone modifications during germ cell development.<\/jats:p>","DOI":"10.3390\/cells15171577","type":"journal-article","created":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T02:08:58Z","timestamp":1788142138000},"page":"1577","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["MBD3 Knockdown Promotes the Differentiation of Bovine iPSCs into Primordial Germ Cell-like Cells Associated with Altered Bivalent Modification of TGF-\u03b2-Related Genes"],"prefix":"10.3390","volume":"15","author":[{"given":"Wen","family":"Yuan","sequence":"first","affiliation":[{"name":"State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, College of Life Science, Inner Mongolia University, Hohhot 010021, China"},{"name":"College of Life Science and 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China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fang","family":"Liu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, College of Life Science, Inner Mongolia University, Hohhot 010021, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoxiong","family":"Geng","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, College of Life Science, Inner Mongolia University, Hohhot 010021, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongli","family":"Yue","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, College of Life Science, Inner Mongolia University, Hohhot 010021, 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J."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"178","DOI":"10.1016\/j.stem.2015.06.014","article-title":"Robust In Vitro Induction of Human Germ Cell Fate from Pluripotent Stem Cells","volume":"17","author":"Sasaki","year":"2015","journal-title":"Cell Stem Cell"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1038\/nmeth.3317","article-title":"HISAT: A fast spliced aligner with low memory requirements","volume":"12","author":"Kim","year":"2015","journal-title":"Nat. Methods"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1093\/bioinformatics\/btp616","article-title":"edgeR: A Bioconductor package for differential expression analysis of digital gene expression data","volume":"26","author":"Robinson","year":"2010","journal-title":"Bioinformatics"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1089\/omi.2011.0118","article-title":"clusterProfiler: An R package for comparing biological themes among gene clusters","volume":"16","author":"Yu","year":"2012","journal-title":"OMICS J. Integr. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1754","DOI":"10.1093\/bioinformatics\/btp324","article-title":"Fast and accurate short read alignment with Burrows-Wheeler transform","volume":"25","author":"Li","year":"2009","journal-title":"Bioinformatics"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"R137","DOI":"10.1186\/gb-2008-9-9-r137","article-title":"Model-based analysis of ChIP-Seq (MACS)","volume":"9","author":"Zhang","year":"2008","journal-title":"Genome Biol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2382","DOI":"10.1093\/bioinformatics\/btv145","article-title":"ChIPseeker: An R\/Bioconductor package for ChIP peak annotation, comparison and visualization","volume":"31","author":"Yu","year":"2015","journal-title":"Bioinformatics"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"111587","DOI":"10.1016\/j.celrep.2022.111587","article-title":"Integration of single-cell transcriptome and chromatin accessibility of early gonads development among goats, pigs, macaques, and humans","volume":"41","author":"Chen","year":"2022","journal-title":"Cell Rep."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1530\/REP-20-0313","article-title":"Similarities between bovine and human germline development revealed by single-cell RNA sequencing","volume":"161","author":"Soto","year":"2021","journal-title":"Reproduction"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"122","DOI":"10.12688\/wellcomeopenres.15381.1","article-title":"Testing the role of SOX15 in human primordial germ cell fate","volume":"4","author":"Sybirna","year":"2019","journal-title":"Wellcome Open Res."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Yuan, W., Wang, J., Long, S., Liu, F., Geng, X., Su, M., Wang, Y., Li, C., Yue, Y., and Li, X. (2025). Knockdown of MBD3 Promotes Pluripotency and In Vitro Differentiation Ability of Bovine iPSCs by Reducing DNA Damage via HIF1A During Reprogramming. Preprints.","DOI":"10.20944\/preprints202501.2005.v1"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"e72811","DOI":"10.7554\/eLife.72811","article-title":"Efficient differentiation of human primordial germ cells through geometric control reveals a key role for Nodal signaling","volume":"11","author":"Jo","year":"2022","journal-title":"eLife"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"517","DOI":"10.1016\/j.stem.2017.09.005","article-title":"Evolutionarily Distinctive Transcriptional and Signaling Programs Drive Human Germ Cell Lineage Specification from Pluripotent Stem Cells","volume":"21","author":"Kojima","year":"2017","journal-title":"Cell Stem Cell"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1038\/s41586-018-0581-5","article-title":"OTX2 restricts entry to the mouse germline","volume":"562","author":"Zhang","year":"2018","journal-title":"Nature"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1437","DOI":"10.1016\/j.cell.2015.05.015","article-title":"The Transcriptome and DNA Methylome Landscapes of Human Primordial Germ Cells","volume":"161","author":"Guo","year":"2015","journal-title":"Cell"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"102003","DOI":"10.1016\/j.isci.2020.102003","article-title":"Human pluripotent stem cell-derived DDX4 and KRT-8 positive cells participate in ovarian follicle-like structure formation","volume":"24","author":"Yu","year":"2021","journal-title":"iScience"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"416","DOI":"10.1038\/nature22812","article-title":"Principles of early human development and germ cell program from conserved model systems","volume":"546","author":"Kobayashi","year":"2017","journal-title":"Nature"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"a022137","DOI":"10.1101\/cshperspect.a022137","article-title":"Signaling Cross Talk between TGF-\u03b2\/Smad and Other Signaling Pathways","volume":"9","author":"Luo","year":"2017","journal-title":"Cold Spring Harb. Perspect. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1530\/rep.1.01075","article-title":"All in the family: TGF-beta family action in testis development","volume":"132","author":"Itman","year":"2006","journal-title":"Reproduction"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"dev201102","DOI":"10.1242\/dev.201102","article-title":"The journey of a generation: Advances and promises in the study of primordial germ cell migration","volume":"151","author":"Barton","year":"2024","journal-title":"Development"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.semcdb.2015.10.029","article-title":"TGF-\u03b2 superfamily signaling in testis formation and early male germline development","volume":"45","author":"Young","year":"2015","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"43851","DOI":"10.1038\/srep43851","article-title":"CD61 promotes the differentiation of canine ADMSCs into PGC-like cells through modulation of TGF-\u03b2 signaling","volume":"7","author":"Fang","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1016\/j.ydbio.2005.05.019","article-title":"Altered primordial germ cell migration in the absence of transforming growth factor beta signaling via ALK5","volume":"284","author":"Carvalho","year":"2005","journal-title":"Dev. Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"4014","DOI":"10.1111\/jcmm.16366","article-title":"ARL4C might serve as a prognostic factor and a novel therapeutic target for gastric cancer: Bioinformatics analyses and biological experiments","volume":"25","author":"Xie","year":"2021","journal-title":"J. Cell. Mol. Med."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Santiago, J.A., Quinn, J.P., and Potashkin, J.A. (2023). Co-Expression Network Analysis Identifies Molecular Determinants of Loneliness Associated with Neuropsychiatric and Neurodegenerative Diseases. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24065909"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"20105","DOI":"10.1074\/jbc.M115.655340","article-title":"Lysosomal-associated Transmembrane Protein 4B (LAPTM4B) Decreases Transforming Growth Factor \u03b21 (TGF-\u03b21) Production in Human Regulatory T Cells","volume":"290","author":"Huygens","year":"2015","journal-title":"J. Biol. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"216902","DOI":"10.1016\/j.canlet.2024.216902","article-title":"Direct contact between tumor cells and platelets initiates a FAK-dependent F3\/TGF-\u03b2 positive feedback loop that promotes tumor progression and EMT in osteosarcoma","volume":"591","author":"Shi","year":"2024","journal-title":"Cancer Lett."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/j.cell.2013.08.056","article-title":"Dnmt3L antagonizes DNA methylation at bivalent promoters and favors DNA methylation at gene bodies in ESCs","volume":"155","author":"Neri","year":"2013","journal-title":"Cell"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"696","DOI":"10.1038\/s41594-020-0443-3","article-title":"Epigenetic priming by Dppa2 and 4 in pluripotency facilitates multi-lineage commitment","volume":"27","author":"Parry","year":"2020","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Fang, S., Wang, J., Liu, G., Qu, B., Chunyu, J., Xu, W., Xiang, J., and Li, X. (2024). DPPA2\/4 Promote the Pluripotency and Proliferation of Bovine Extended Pluripotent Stem Cells by Upregulating the PI3K\/AKT\/GSK3\u03b2\/\u03b2-Catenin Signaling Pathway. Cells, 13.","DOI":"10.3390\/cells13050382"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1041","DOI":"10.1002\/stem.652","article-title":"Differential recruitment of methyl CpG-binding domain factors and DNA methyltransferases by the orphan receptor germ cell nuclear factor initiates the repression and silencing of Oct4","volume":"29","author":"Gu","year":"2011","journal-title":"Stem Cells"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1038\/nature09607","article-title":"c-Jun N-terminal phosphorylation antagonises recruitment of the Mbd3\/NuRD repressor complex","volume":"469","author":"Aguilera","year":"2011","journal-title":"Nature"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"e2104841118","DOI":"10.1073\/pnas.2104841118","article-title":"AP-1 is a temporally regulated dual gatekeeper of reprogramming to pluripotency","volume":"118","author":"Markov","year":"2021","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"3831","DOI":"10.1093\/nar\/gkf509","article-title":"Dnmt3L is a transcriptional repressor that recruits histone deacetylase","volume":"30","author":"Deplus","year":"2002","journal-title":"Nucleic Acids Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1530\/REP-15-0031","article-title":"Dnmt3l-knockout donor cells improve somatic cell nuclear transfer reprogramming efficiency","volume":"150","author":"Liao","year":"2015","journal-title":"Reproduction"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"10680","DOI":"10.1128\/JVI.01176-14","article-title":"Ectopic DNMT3L triggers assembly of a repressive complex for retroviral silencing in somatic cells","volume":"88","author":"Kao","year":"2014","journal-title":"J. Virol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1038\/nature12587","article-title":"Deterministic direct reprogramming of somatic cells to pluripotency","volume":"502","author":"Rais","year":"2013","journal-title":"Nature"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1262\/jrd.2023-087","article-title":"Efficient derivation of embryonic stem cells and primordial germ cell-like cells in cattle","volume":"70","author":"Shirasawa","year":"2024","journal-title":"J. Reprod. Dev."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"e202000974","DOI":"10.26508\/lsa.202000974","article-title":"GATA transcription factors, SOX17 and TFAP2C, drive the human germ-cell specification program","volume":"4","author":"Kojima","year":"2021","journal-title":"Life Sci. Alliance"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"16061","DOI":"10.1073\/pnas.1315204110","article-title":"A set of genes critical to development is epigenetically poised in mouse germ cells from fetal stages through completion of meiosis","volume":"110","author":"Lesch","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1777","DOI":"10.1016\/j.celrep.2013.04.032","article-title":"Bivalent chromatin marks developmental regulatory genes in the mouse embryonic germline in vivo","volume":"3","author":"Sachs","year":"2013","journal-title":"Cell Rep."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1038\/nature19362","article-title":"Distinct features of H3K4me3 and H3K27me3 chromatin domains in pre-implantation embryos","volume":"537","author":"Liu","year":"2016","journal-title":"Nature"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1038\/s41421-023-00519-1","article-title":"Resetting histone modifications during human prenatal germline development","volume":"9","author":"Gao","year":"2023","journal-title":"Cell Discov."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1038\/s41594-020-0453-1","article-title":"Dppa2 and Dppa4 safeguard bivalent chromatin in order to establish a pluripotent epigenome","volume":"27","author":"Lim","year":"2020","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"2844","DOI":"10.1016\/j.stemcr.2021.10.008","article-title":"DPPA2, DPPA4, and other DPPA factor epigenomic functions in cell fate and cancer","volume":"16","author":"Klein","year":"2021","journal-title":"Stem Cell Rep."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/15\/17\/1577\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T02:27:32Z","timestamp":1788143252000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/15\/17\/1577"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,30]]},"references-count":46,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["cells15171577"],"URL":"https:\/\/doi.org\/10.3390\/cells15171577","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,30]]}}}