{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,28]],"date-time":"2026-08-28T15:52:40Z","timestamp":1787932360822,"version":"build-2784847793"},"reference-count":21,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2025,2,6]],"date-time":"2025-02-06T00:00:00Z","timestamp":1738800000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Gratama-stichting","award":["2019-14"],"award-info":[{"award-number":["2019-14"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Background: In prenatal genetic diagnostics, the detection of single-gene defects relies on chorionic villus sampling (CVS) and amniocentesis, which carry a miscarriage risk of 0.2\u20130.3%. To mitigate this risk, fetal trophoblasts have been isolated from a Papanicolaou smear using Trophoblast Retrieval and Isolation from the Cervix (TRIC). However, this method is labor-intensive and has been shown to be challenging to implement in clinical practice. Here, we describe our experiences in using semi-automated immunomagnetic cell sorting for isolating trophoblasts from clinically obtained Papanicolaou smears during ongoing pregnancies. Methods: Using HLA-G-positive Jeg-3 and HLA-G-negative HeLa cell lines in 10%, 1%, and 0.1% dilutions, we tested and optimized the isolation of HLA-G-positive cells using FACS and semi-automated immunomagnetic cell sorting. We used the latter technique for isolation of HLA-G-positive cells from Papanicolaou smears collected from 26 pregnant women, gestational age between 6 and 20 weeks, who underwent CVS. Results: In four independent dilution series, the mean percentages of Jeg-3 cells went from 7.1% to 53.5%, 0.9% to 32.6%, and 0.4% to 2.6% (7.5, 36, and 6.5-fold enrichment, respectively) using immunomagnetic cell sorting. After sorting of the Papanicolaou smears, HLA-G-positive cells were moderately increased in the positive (14.61 vs. 11.63%) and decreased in the negative fraction (7.87 vs. 11.63%) compared to baseline pre-sorting. However, we could not identify fetal cells using XY-chromosomal FISH in a male sample. Conclusions: Our study supports previous findings that careful sampling of fetal cells from Papanicolaou smears in a clinical context poses significant challenges to cell retrieval.<\/jats:p>","DOI":"10.3390\/diagnostics15030386","type":"journal-article","created":{"date-parts":[[2025,2,6]],"date-time":"2025-02-06T06:30:29Z","timestamp":1738823429000},"page":"386","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Limitations of Semi-Automated Immunomagnetic Separation of HLA-G-Positive Trophoblasts from Papanicolaou Smears for Prenatal Genetic Diagnostics"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7138-8642","authenticated-orcid":false,"given":"Eddy N.","family":"de Boer","sequence":"first","affiliation":[{"name":"Department of Genetics, University Medical Center Groningen, University of Groningen, 9700 RB Groningen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicole","family":"Corsten-Janssen","sequence":"additional","affiliation":[{"name":"Department of Genetics, University Medical Center Groningen, University of Groningen, 9700 RB Groningen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elles","family":"Wierenga","sequence":"additional","affiliation":[{"name":"Department of Genetics, University Medical Center Groningen, University of Groningen, 9700 RB Groningen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1522-9137","authenticated-orcid":false,"given":"Theo","family":"Bijma","sequence":"additional","affiliation":[{"name":"Flow Cytometry Unit, Pathology & Medical Biology, University Medical Center Groningen, University of Groningen, 9700 RB Groningen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-1907-8697","authenticated-orcid":false,"given":"Jurjen T.","family":"Knapper","sequence":"additional","affiliation":[{"name":"Department of Genetics, University Medical Center Groningen, University of Groningen, 9700 RB Groningen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5615-8304","authenticated-orcid":false,"given":"Gerard J. te","family":"Meerman","sequence":"additional","affiliation":[{"name":"Department of Genetics, University Medical Center Groningen, University of Groningen, 9700 RB Groningen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gwendolyn T. R.","family":"Manten","sequence":"additional","affiliation":[{"name":"Department of Obstetrics, Isala Clinic, 8025 AB Zwolle, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-2916-1524","authenticated-orcid":false,"given":"Nine V. A. M.","family":"Knoers","sequence":"additional","affiliation":[{"name":"Department of Genetics, University Medical Center Groningen, University of Groningen, 9700 RB Groningen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9253-8200","authenticated-orcid":false,"given":"Katelijne","family":"Bouman","sequence":"additional","affiliation":[{"name":"Department of Genetics, University Medical Center Groningen, University of Groningen, 9700 RB Groningen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leonie K.","family":"Duin","sequence":"additional","affiliation":[{"name":"Department of Obstetrics, University Medical Center Groningen, University of Groningen, 9700 RB Groningen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9283-2207","authenticated-orcid":false,"given":"Cleo C. van","family":"Diemen","sequence":"additional","affiliation":[{"name":"Department of Genetics, University Medical Center Groningen, University of Groningen, 9700 RB Groningen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,6]]},"reference":[{"key":"ref_1","first-page":"557","article-title":"March of Dimes releases premature birth report card: Kentucky receives \u201cF\u2019\u2019\u2019\u2014KMA joins healthy babies coalition","volume":"106","author":"Christianson","year":"2008","journal-title":"J. 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