HLA‐G positive trophoblastic cells in transcervical samples and their isolation and analysis by laser microdissection and QF‐PCR

Bulmer, Judith N.; Cioni, Riccardo; Bussani, Cecilia; Cirigliano, Vincenzo; Solé, Françesc; Costa, Carlota; García, P. Peña; Adinolfí, Matteo

doi:10.1002/pd.511

Cited by 33 publications

(29 citation statements)

References 27 publications

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“…In 2003, Bulmer et al employed MAbs against HLA-G to identify cytotrophoblasts cells in TCC samples retrieved by IUL (21). Cytotrophoblast cells characterized by their large, irregular hyperchromatic nuclei were HLA-G positive and were identified in 12 of 23 (52%) TCC samples.…”

Section: Methods Of Analysismentioning

confidence: 99%

“…It should also be simple to perform and cost effective, with minimal inter-observer variability. A number of techniques have been devised to retrieve TCC samples from the endocervical canal and the endometrial cavity, including smears obtained with cotton swabs or a cytobrush (14–16), aspiration of cervical mucus with a catheter (17–19), endometrial biopsy with a Pipelle (20), and lavage of the endocervical canal (21–24) or the uterine cavity (14;22;25), all with variable success (Table 1). …”

Section: Retrieval Of Trophoblast Cells From the Cervixmentioning

confidence: 99%

“…Similarly, Fejgin et al reported good results without using mucolytics (15). Others (21;30;31) reported using mucolytics only when the mucus was thick without affecting the specimen quality. In still another study, Cioni and colleagues (26) compared the TCC specimens collected by cytobrush that were processed using N -acetylcysteine, dithiothreitol, or no mucolytic, and reported neither to be superior.…”

Section: Post-collection Processingmentioning

confidence: 99%

See 2 more Smart Citations

Transcervical retrieval of fetal cells in the practice of modern medicine: a review of the current literature and future direction

Imudia

Kumar

Diamond

et al. 2010

Fertility and Sterility

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Objective-To review published methods for transcervical collection of fetal cells and to assess the potential of this approach for application in prenatal diagnosis.Design-Retrospective analysis of efforts at prenatal diagnosis using trophoblast cells shed into the lower uterine pole that accumulate within the cervical mucus at the level of the internal os.Results-Minimally invasive techniques that include cervical mucus aspiration, cervical swabbing, and cervical or intrauterine lavage can be used to retrieve trophoblast cells during the first trimester for diagnostic purposes, including prenatal genetic analysis. Fetal cells have been identified in these specimens with success rates that vary from 40 to 90%. The disparity in reported success rates can be a function of gestational age, collection method, operator variability, detection sensitivity, or pregnancy status. Molecular approaches have been devised to determine fetal sex and identify aneuploidies. Antibody markers have proven useful to select trophoblast cells for genetic analysis and to demonstrate that the abundance of recoverable fetal cells diminishes in abnormal gestations, such as ectopic pregnancy and blighted ovum.Conclusions-Transcervical collection of fetal cells offers several avenues for prenatal diagnosis that with further refinement could one day provide valuable information for the management of ongoing pregnancies.

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Section: Methods Of Analysismentioning

confidence: 99%

Section: Retrieval Of Trophoblast Cells From the Cervixmentioning

confidence: 99%

Section: Post-collection Processingmentioning

confidence: 99%

See 1 more Smart Citation

Transcervical retrieval of fetal cells in the practice of modern medicine: a review of the current literature and future direction

Imudia

Kumar

Diamond

et al. 2010

Fertility and Sterility

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“…As such, there has been considerable interest in refining non-invasive methods of PND, which can be conducted in early pregnancy. In recent years, several groups including our own have shown considerable success in isolating fetal cells from the cervical mucous for use in PND Bussani et al, 2002;Bulmer et al, 2003;Katz-Jaffe et al, 2004]. Eventually, these methods of non-invasive PND will be developed to a high level of reliability, enabling small numbers of fetal cells to be retrieved for genetic analysis.…”

Section: Introductionmentioning

confidence: 98%

A QF-PCR system to detect chromosome 13 aneuploidy from as few as ten cells

2005

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Complete and partial trisomies of chromosome 13 are characterized by abnormal fetal development and birth defects. Despite the severe abnormalities associated with trisomy 13, some couples elect not to undergo invasive prenatal diagnosis (PND) due to the 0.5%-1.0% risk of pregnancy loss. As a result, current studies are focusing on refining non-invasive prenatal diagnostic techniques, such as screening fetal cells isolated from maternal blood or cervical smears. As these techniques only provide a limited number of cells for analysis, any progress in this field depends on the development of a highly sensitive genetic screening strategy. We have developed a quantitative fluorescent PCR (QF-PCR) system capable of detecting chromosome 13 aneuploidy from as few as 10 cells. The system was further validated by screening 13 amniocyte samples, three of which had been diagnosed by FISH as having chromosomal abnormalities involving chromosome 13. In all cases, the QF-PCR results were concordant with those obtained using FISH. The high reliability (99%) and accuracy (96%) of the QF-PCR system at the 10 cell level makes this technique ideal for use in non-invasive PND.

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“…A more recent study using PCR and FISH analyses on transcervical cells resulted in poor detection of fetal cells ]. To distinguish trophoblast cells from the predominant maternal cell population in transcervical cell samples, antibodies directed against placental antigens were employed [Koumantaki et al, 2001;Bulmer et al, 2003]. These analyses resulted in an overall detection rate of trophoblasts of 25 to 93%.…”

Section: Prenatal Detection Of Down Syndrome Through Detection Of Tromentioning

confidence: 99%

Non Invasive Prenatal Diagnosis of Down Syndrome

Kappou¹,

Papadopoulou²,

Sifakis³

2011

Prenatal Diagnosis and Screening for Down Syndrome

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HLA‐G positive trophoblastic cells in transcervical samples and their isolation and analysis by laser microdissection and QF‐PCR

Abstract: These results confirm that, during an early phase of gestation, fetal cells are released in the lower uterine cavity and that they can be isolated and analysed for prenatal diagnosis of single gene defects and aneuploidies.

Cited by 33 publications

References 27 publications

Transcervical retrieval of fetal cells in the practice of modern medicine: a review of the current literature and future direction

Transcervical retrieval of fetal cells in the practice of modern medicine: a review of the current literature and future direction

A QF-PCR system to detect chromosome 13 aneuploidy from as few as ten cells

Non Invasive Prenatal Diagnosis of Down Syndrome

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