{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T13:31:21Z","timestamp":1785159081857,"version":"3.55.0"},"reference-count":39,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,5,9]],"date-time":"2025-05-09T00:00:00Z","timestamp":1746748800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005039","name":"Leids Universitair Medisch Centrum","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100005039","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Maternal-fetal HLA compatibility influences pregnancy outcome, including preeclampsia risk. Cervical extravillous trophoblasts (EVT) in early pregnancy provide a non-invasive source for fetal genome acquisition, potentially enabling fetal HLA typing for obstetric risk assessment. This study aimed to achieve fetal HLA typing through EVT isolation using HLA-G-coupled nanoparticle immunomagnetic separation (TRIC) and fluorescence-activated cell sorting (FACS).<\/jats:p><\/jats:sec><jats:sec><jats:title>Method<\/jats:title><jats:p>Cervical samples from 32 pregnant women were collected by cytobrush. Saliva and umbilical cord blood (n=13) served as maternal and fetal HLA genotype controls, respectively. Cervical samples from non-pregnant women, primary cultured EVT, and cryo-sectioned term placentas served as controls for cell phenotype, protein expression, and effect of fixation. FACS and TRIC were applied to isolate EVT from maternal cells, followed by RSSO-PCR for HLA typing. EVT presence pre- and post-isolation was determined through HLA-G, \u03b2-hCG, and Cytokeratin-7 (CK-7) expression. TRIC was optimized by improving antibody-binding-efficiency, and comparing three (nano)beads types and two magnets.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Purity and yield of HLA-G<jats:sup>+<\/jats:sup>\u03b2-hCG<jats:sup>+<\/jats:sup>CK-7<jats:sup>+<\/jats:sup> cells after TRIC failed to match pre-isolation HLA-G<jats:sup>+<\/jats:sup> cell counts, despite protocol optimization. FACS revealed a fetal HLA genotype. In contrast, only the maternal HLA genotype was detected in TRIC-isolated cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>EVT counts and maternal cell contamination limit reliable fetal HLA typing from cervical samples. Refining non-invasive EVT isolation techniques may enable fetal HLA typing to be included in risk assessment of pregnancy complications.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2025.1575086","type":"journal-article","created":{"date-parts":[[2025,5,9]],"date-time":"2025-05-09T04:10:39Z","timestamp":1746763839000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":3,"title":["Cervical EVT isolation for non-invasive fetal HLA typing in early pregnancy is limited by purity and maternal cell contamination; a methodological comparison"],"prefix":"10.3389","volume":"16","author":[{"given":"Liseanne J.","family":"van \u2018t Hof","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johanna M.","family":"Kapsenberg","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos J.M.","family":"Drabbels","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lotte E.","family":"van der Meeren","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dave L.","family":"Roelen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Eikmans","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie-Louise P.","family":"van der Hoorn","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,5,9]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1038\/nm.3160","article-title":"Fetomaternal immune cross-talk and its consequences for maternal and offspring\u2019s health","volume":"19","author":"Arck","year":"2013","journal-title":"Nat Medicine"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1016\/j.humimm.2015.02.004","article-title":"HLA associations and HLA sharing in recurrent miscarriage: A systematic review and meta-analysis","volume":"76","author":"Meuleman","year":"2015","journal-title":"Hum Immunol"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.5935\/1518-0557.20180001","article-title":"Is the risk of preeclampsia higher in donor oocyte pregnancies? 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