Evidence for Feasibility of Fetal Trophoblastic Cell-Based Noninvasive Prenatal Testing

Breman, Amy M.; Chow, Jennifer; U'Ren, Lance; Normand, Elizabeth A.; Qdaisat, Sadeem; Zhao, Lei; Henke, David; Chen, Rui; Shaw, Chad A.; Jackson, Laird G.; Yang, Yaping; Vossaert, Liesbeth; Needham, Rachel H. V.; Chang, Elizabeth; Campton, Daniel; Werbin, Jeffrey L.; Seubert, Ron C.; Veyver, Ignatia B. Van den; Stilwell, Jackie L.; Kaldjian, Eric P.; Beaudet, Arthur L.

doi:10.1097/01.ogx.0000511968.79993.bf

Cited by 22 publications

(48 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…(A method for preparation and single-cell analysis of live cells using chamber slides has been developed but is not discussed here.) Prepared DNA may be used for various types of analysis including PCR, mass spectrometry, comparative genomic hybridization, and nextgeneration sequencing (12,15,17). Specific gene sequencing of identified CTCs has confirmed mutations present in tumor tissue samples.…”

Section: Single Cell Retrieval Systemmentioning

confidence: 99%

“…Using similar methods of sample preparation, multiparameter staining, imaging and analysis, and cell retrieval, the platform has been applied to other rare cell investigations. Areas include fetal cell analysis for noninvasive prenatal genetic testing (17), minimal residual disease monitoring of liquid tumors, identification of rare immune cell subsets in blood, and pathological analysis of fine needle aspirate, other cytology samples, and tissue sections.…”

Section: Application Beyond Ctcsmentioning

confidence: 99%

See 1 more Smart Citation

The RareCyte® platform for next‐generation analysis of circulating tumor cells

et al. 2018

Self Cite

View full text Add to dashboard Cite

Circulating tumor cells (CTCs) can reliably be identified in cancer patients and are associated with clinical outcome. Next‐generation “liquid biopsy” technologies will expand CTC diagnostic investigation to include phenotypic characterization and single‐cell molecular analysis. We describe here a rare cell analysis platform designed to comprehensively collect and identify CTCs, enable multi‐parameter assessment of individual CTCs, and retrieve single cells for molecular analysis. The platform has the following four integrated components: 1) density‐based separation of the CTC‐containing blood fraction and sample deposition onto microscope slides; 2) automated multiparameter fluorescence staining; 3) image scanning, analysis, and review; and 4) mechanical CTC retrieval. The open platform utilizes six fluorescence channels, of which four channels are used to identify CTC and two channels are available for investigational biomarkers; a prototype assay that allows three investigational biomarker channels has been developed. Single‐cell retrieval from fixed slides is compatible with whole genome amplification methods for genomic analysis. © 2018 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry.

show abstract

Section: Single Cell Retrieval Systemmentioning

confidence: 99%

Section: Application Beyond Ctcsmentioning

confidence: 99%

The RareCyte® platform for next‐generation analysis of circulating tumor cells

et al. 2018

Self Cite

View full text Add to dashboard Cite

show abstract

“…However, using circulating fetal cells for prenatal diagnosis compared with cffDNA allows for whole genome analysis on DNA from pure fetal cells without maternal contamination 7‐9 . Several different types of circulating fetal cells have been identified and targeted for prenatal diagnosis, including lymphoid progenitor cells, nucleated red blood cells and fEVTs 10‐14 …”

Section: Introductionmentioning

confidence: 99%

Do fetal extravillous trophoblasts circulate in maternal blood postpartum?

Looij

Singh²,

Hatt³

et al. 2020

Acta Obstet Gynecol Scand

View full text Add to dashboard Cite

This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made. AbstractIntroduction: Circulating fetal extravillous trophoblasts may offer a superior alternative to cell-free fetal DNA for noninvasive prenatal testing. Cells of fetal origin are a pure source of fetal genome; hence, unlike the cell-free noninvasive prenatal test, the fetal cell-based noninvasive prenatal test is not expected to be affected by maternal DNA. However, circulating fetal cells from previous pregnancies may lead to confounding results. Material and methods:To study whether fetal trophoblast cells persist in maternal circulation postpartum, blood samples were collected from 11 women who had given birth to a boy, with blood sampling at 1-3 days (W0), 4-5 weeks (W4-5), around 8 weeks (W8) and around 12 weeks (W12) postpartum. The existence of fetal extravillous trophoblasts was verified either by X and Y chromosome fluorescence in situ hybridization analysis or by short tandem repeat analysis. To exclude technological bias in isolating fetal cells, blood samples were also collected from 10 pregnant women between a gestational age of 10 and 14 weeks, the optimal time frame for cell-based noninvasive prenatal test sampling. All the samples were processed according to protocols established by ARCEDI Biotech for fetal extravillous trophoblast enrichment and isolation.Results: Fetal extravillous trophoblasts were found in all the 10 samples from pregnant women between a gestational age of 10 and 14 weeks. However, only 4 of 11 blood samples taken from women at 1-3 days postpartum rendered fetal extravillous trophoblasts, and only 2 of 11 samples rendered fetal extravillous trophoblasts at 4 weeks postpartum. Conclusions:In this preliminary dataset on few pregnancies, none of the samples rendered any fetal cells at or after 8 weeks postpartum, showing that cell-based noninvasive prenatal testing based on fetal extravillous trophoblasts is unlikely to be influenced by circulating cells from previous pregnancies. K E Y W O R D S chorionic villi sampling, fetal cells, fetal extravillous trophoblasts, noninvasive prenatal testing, prenatal diagnosis S U PP O RTI N G I N FO R M ATI O N Additional supporting information may be found online in the Supporting Information section.

show abstract

“…As expected, each of these conditions has been associated with increased NRBCs in the newborn 9 . The other objective is to screen and extract fNRBCs from maternal peripheral blood for non-invasive prenatal diagnosis (NIPD) 10–12 . The choice of fNRBCs as ideal target cells is based on the following parameters 13,14 : (1) presence of intact nuclei containing the complete fetal genome in fNRBCs, which is a prerequisite for prenatal analysis; (2) limited life span of fNRBCs in the maternal circulation, which can be differentiated morphologically from maternal cells; and (3) presence of distinct cell markers, such as epsilon hemoglobin transferrin receptor (CD71) 15 , thrombospondin receptor (CD36), and glycophorin A (GPA) in fNRBCs that enable isolation of these rare cells from large volumes of maternal blood.…”

Section: Introductionmentioning

confidence: 99%

Deep learning-based adaptive detection of fetal nucleated red blood cells

Sun

Wang

Zhao

et al. 2020

Preprint

View full text Add to dashboard Cite

15Aim: this study, we established an artificial intelligence system for rapid 16 identification of fetal nucleated red blood cells (fNRBCs). 17Method: Density gradient centrifugation and magnetic-activated cell sorting were 18 used for the separation of fNRBCs from umbilical cord blood. The cell block 19 technique was used for fixation. We proposed a novel preprocessing method based on 20 imaging characteristics of fNRBCs for region of interest (ROI) extraction, which 21 automatically segmented individual cells in peripheral blood cell smears. The 22 discriminant information from ROIs was encoded into a feature vector and 23 pathological diagnosis were provided by the prediction network. 24Results: Four umbilical cord blood samples were collected and validated based on a 25 large dataset containing 260 samples. Finally, the dataset was classified into 3,720 and 26 1,040 slides for training and testing, respectively. In the test set, classifier obtained 27 98.5% accuracy and 96.5% sensitivity. 28Conclusion: Therefore, this study offers an effective and accurate method for 29 fNRBCs preservation and identification. 30 Keywords: fetal nucleated red blood cells, cell-block, deep learning, non-invasive 31 prenatal diagnosis 32 Introduction 33The clinical application of fNRBCs during pregnancy could be classified into two 34 main categories 1,2 . One is the prognosis of possible diseases in pregnant women by 35 counting fNRBCs in umbilical cord blood. Chronic tissue hypoxia results in increased 36 levels of erythropoietin, which, in turn, leads to stimulation of erythropoiesis and 37 increased numbers of circulating nucleated red blood cells (NRBCs) 1,3-5 . Increased 38 umbilical cord levels of erythropoietin have been reported in pregnancies complicated 39 by intrauterine growth restriction, maternal hypertension, preeclampsia, maternal 40 smoking, Rh isoimmunization, and maternal diabetes 5-8 . As expected, each of these 41 conditions has been associated with increased NRBCs in the newborn 9 . The other 42 objective is to screen and extract fNRBCs from maternal peripheral blood for 43 non-invasive prenatal diagnosis (NIPD) 10-12 . The choice of fNRBCs as ideal target 44 cells is based on the following parameters 13,14 : (1) presence of intact nuclei containing 45 the complete fetal genome in fNRBCs, which is a prerequisite for prenatal analysis; (2) 46 limited life span of fNRBCs in the maternal circulation, which can be differentiated 47 55 cells) 16,17 . Several fNRBC enrichment methods based on different principles have 56 been reported, such as density gradient centrifugation (DGC) 13,18 , 57 fluorescence-activated cell sorting (FACS) 19 , and magnetic-activated cell sorting 58 (MACS) 20 , dielectrophoresis, and microfluidics based technologies 21,22 . Nevertheless, 59 long-term preservation of samples and rapid identification of target cells (fNRBCs) 60 still present considerable challenges. 61 Since the identification of fNRBCs in large number of cell block slices represent a 62 huge manual burden on p...

show abstract

Evidence for Feasibility of Fetal Trophoblastic Cell-Based Noninvasive Prenatal Testing

Cited by 22 publications

References 0 publications

The RareCyte® platform for next‐generation analysis of circulating tumor cells

The RareCyte® platform for next‐generation analysis of circulating tumor cells

Do fetal extravillous trophoblasts circulate in maternal blood postpartum?

Deep learning-based adaptive detection of fetal nucleated red blood cells

Contact Info

Product

Resources

About