Rarity of fetal cells in exocervical samples for noninvasive prenatal diagnosis

Laura Bourlard; Yannick Manigart; Catherine Donner; Guillaume Smits; Julie Désir; Isabelle Migeotte; Bruno Pichon

doi:10.1515/jpm-2021-0291

Article

Rarity of fetal cells in exocervical samples for noninvasive prenatal diagnosis

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Published/Copyright: December 31, 2021

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From the journal Journal of Perinatal Medicine Volume 50 Issue 4

Abstract

Objectives

The possibility to isolate fetal cells from pregnant women cervical samples has been discussed for five decades but is not currently applied in clinical practice. This study aimed at offering prenatal genetic diagnosis from fetal cells obtained through noninvasive exocervical sampling and immuno-sorted based on expression of HLA-G.

Methods

We first developed and validated robust protocols for cell detection and isolation on control cell lines expressing (JEG-3) or not (JAR) the HLA-G antigen, a specific marker for extravillous trophoblasts. We then applied these protocols to noninvasive exocervical samples collected from pregnant women between 6 and 14 weeks of gestational age. Sampling was performed through insertion and rotation of a brush at the ectocervix close to the external os of the endocervical canal. Finally, we attempted to detect and quantify trophoblasts in exocervical samples from pregnant women by ddPCR targeting the male SRY locus.

Results

For immunohistochemistry, a strong specific signal for HLA-G was observed in the positive control cell line and for rare cells in exocervical samples, but only in non-fixative conditions. HLA-G positive cells diluted in HLA-G negative cells were isolated by flow cytometry or magnetic cell sorting. However, no HLA-G positive cells could be recovered from exocervical samples. SRY gene was detected by ddPCR in exocervical samples from male (50%) but also female (27%) pregnancies.

Conclusions

Our data suggest that trophoblasts are too rarely and inconstantly present in noninvasive exocervical samples to be reliably retrieved by standard immunoisolation techniques and therefore cannot replace the current practice for prenatal screening and diagnosis.

Keywords: cervix; fetal cells; noninvasive prenatal diagnosis

Corresponding author: Bruno Pichon, PhD, Center of Human Genetics, Université Libre de Bruxelles – Erasme Hospital, Route de Lennik 808, 1070 Brussels, Belgium, Phone: +003225556417. E-mail: bruno.pichon@erasme.ulb.ac.be

Funding source: The Belgian Kids' Fund for Pediatric Research (BKF)

Award Identifier / Grant number: PhD Fellowship

Funding source: Fondation Paul, Suzanne, Renée Lippens

Award Identifier / Grant number: Grant for research equipment and products

Funding source: Le Fond Erasme pour la recherche médicale http://dx.doi.org/10.13039/100015400

Award Identifier / Grant number: Grant for research equipment and products

Acknowledgments

For author contributions. For source of funding.

Research funding: The Belgian Kids' Fund to finance the student as PhD fellowship. Bourse of Fondation Lippens to set up clean laboratory and buy the equipment. Bourse of Fond Erasme, obtained twice, to buy reagents and material products necessary to achieve study.
Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission. LB performed experiments, analyzed data and wrote the draft of the manuscript. YM and CD provided exocervical samples. GS, JD and IM participated to conceptualization, data analysis, and revised the manuscript. BP conceptualized the study, designed experiments, analyzed data and revised the manuscript.
Competing interests: Authors state no conflict of interest.
Informed consent: Informed consent was obtained from all individuals included in this study.
Ethical approval: The protocol of this present study was approved by the Institutional Ethical Committee. All included patients signed the informed consent after provided appropriate study information.

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Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/jpm-2021-0291).

Received: 2021-06-05

Accepted: 2021-12-04

Published Online: 2021-12-31

Published in Print: 2022-05-25

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Supplementary Material

Articles in the same Issue

https://doi.org/10.1515/jpm-2021-0291

Keywords for this article

cervix; fetal cells; noninvasive prenatal diagnosis