Trophoblasts Isolated from the Maternal Circulation: In Vitro Expansion and Potential Application in Non-invasive Prenatal Diagnosis

2005 ◽  
Vol 53 (3) ◽  
pp. 337-339 ◽  
Author(s):  
Esther Guetta ◽  
Liat Gutstein-Abo ◽  
Gad Barkai
Keyword(s):  
Prenatal Diagnosis ◽  
Maternal Blood ◽  
Male Fetus ◽  
Maternal Circulation ◽  
Fetal Cells ◽  
Non Invasive ◽  
In Vitro Expansion ◽  
Feasible Option

Prenatal diagnosis based on rare fetal cells in maternal blood is currently not a feasible option. An effort was made to improve cell yields by targeting trophoblast cells. After sorting, the HLA-G-positive cell fraction was analyzed directly or after culture. In situ hybridization technology was applied to prove fetal cell source in samples from women carrying a male fetus and to predict gender in samples without previous knowledge of fetal sex. In vitro culture led to a significant increase in fetal cells and accurate gender prediction in 93% of these samples. This approach might be useful for non-invasive prenatal diagnosis.

scholarly journals Development of a Specific Monoclonal Antibody to Detect Male Cells Expressing the RPS4Y1 Protein

2021 ◽  
Vol 22 (4) ◽  
pp. 2001
Author(s):  
Silvia Spena ◽  
Chiara Cordiglieri ◽  
Isabella Garagiola ◽  
Flora Peyvandi
Keyword(s):  
Monoclonal Antibody ◽  
Prenatal Diagnosis ◽  
Maternal Blood ◽  
Limiting Factors ◽  
Inherited Disease ◽  
Specific Monoclonal Antibody ◽  
Native Form ◽  
Fetal Cells ◽  
Non Invasive ◽  
Reliable Isolation

Hemophilia is an X-linked recessive bleeding disorder. In pregnant women carrier of hemophilia, the fetal sex can be determined by non-invasive analysis of fetal DNA circulating in the maternal blood. However, in case of a male fetus, conventional invasive procedures are required for the diagnosis of hemophilia. Fetal cells, circulating in the maternal bloodstream, are an ideal target for a safe non-invasive prenatal diagnosis. Nevertheless, the small number of cells and the lack of specific fetal markers have been the most limiting factors for their isolation. We aimed to develop monoclonal antibodies (mAbs) against the ribosomal protein RPS4Y1 expressed in male cells. By Western blotting, immunoprecipitation and immunofluorescence analyses performed on cell lysates from male human hepatoma (HepG2) and female human embryonic kidney (HEK293) we developed and characterized a specific monoclonal antibody against the native form of the male RPS4Y1 protein that can distinguish male from female cells. The availability of the RPS4Y1-targeting monoclonal antibody should facilitate the development of novel methods for the reliable isolation of male fetal cells from the maternal blood and their future use for non-invasive prenatal diagnosis of X-linked inherited disease such as hemophilia.

563: In vitro culture of trophoblasts isolated from maternal circulation: A potential non-invasive approach to prenatal diagnosis

2008 ◽  
Vol 199 (6) ◽  
pp. S164
Author(s):  
Isabelle Desitter ◽  
Patrick Rozenberg ◽  
Naoual Benali-Furet ◽  
Yvon Cayre ◽  
Bussieres Laurence
Keyword(s):  
Prenatal Diagnosis ◽  
In Vitro Culture ◽  
Maternal Circulation ◽  
Invasive Approach ◽  
Non Invasive

scholarly journals Isolation and Enrichment of Circulating Fetal Cells for NIPD: An Overview

Diagnostics ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 2239
Author(s):  
Giulia Sabbatinelli ◽  
Donatella Fantasia ◽  
Chiara Palka ◽  
Elisena Morizio ◽  
Melissa Alfonsi ◽  
...  
Keyword(s):  
Prenatal Diagnosis ◽  
High Efficiency ◽  
Diagnostic Testing ◽  
Genetic Research ◽  
Clinical Genetics ◽  
Maternal Circulation ◽  
Fetal Cells ◽  
Non Invasive ◽  
Prenatal Diagnostic ◽  
Maternal Peripheral Blood

Prenatal diagnosis plays a crucial role in clinical genetics. Non-invasive prenatal diagnosis using fetal cells circulating in maternal peripheral blood has become the goal of prenatal diagnosis, to obtain complete fetal genetic information and avoid risks to mother and fetus. The development of high-efficiency separation technologies is necessary to obtain the scarce fetal cells from the maternal circulation. Over the years, multiple approaches have been applied, including choice of the ideal cell targets, different cell recovering technologies, and refined cell isolation yield procedures. In order to provide a useful tool and to give insights about limitations and advantages of the technologies available today, we review the genetic research on the creation and validation of non-invasive prenatal diagnostic testing protocols based on the rare and labile circulating fetal cells during pregnancy.

scholarly journals The Fetal Erythroblast Is Not the Optimal Target for Non-invasive Prenatal Diagnosis: Preliminary Results

2005 ◽  
Vol 53 (3) ◽  
pp. 331-336 ◽  
Author(s):  
Steen Kølvraa ◽  
Britta Christensen ◽  
Lene Lykke-Hansen ◽  
John Philip
Keyword(s):  
Prenatal Diagnosis ◽  
Y Chromosome ◽  
Maternal Blood ◽  
Cell Type ◽  
Fetal Cells ◽  
Non Invasive ◽  
A Cell ◽  
Optimal Target ◽  
Automated Scanning ◽  
Independent Marker

Fetal cells, present in the blood of pregnant women, are potential targets for non-invasive prenatal diagnosis. The fetal erythroblast has been the favorite target cell type. We investigated four methods of enrichment for fetal erythroblasts, identifying only three fetal erythroblasts in 573 ml of maternal blood. This is much less than the expected two to six fetal cells per ml of maternal blood. Hamada and Krabchi used a cell type-independent marker, i.e., the Y chromosome in maternal blood from male pregnancies after Carnoy fixation, leaving the nuclei for hybridization with X-and Y-chromosome-specific probes. We found with a similar technique 28 fetal cells in 15 ml of maternal blood. The fetal origin of cells was confirmed by hybridizing the nuclei with X- and Y-chromosome-specific probes, using two consecutive hybridizations with the two probes in opposite colors (reverse FISH). Candidate fetal cells were inspected after each hybridization. Only cells that were found to change the color of both probe signals from first to second hybridization were diagnosed as fetal. To reduce the labor-intensive slide screening load, we used semi-automated scanning microscopy to search for candidate cells. We conclude that erythroblasts form only a small fraction of fetal cells present in maternal blood.

HARVESTING FETAL CELLS FROM THE MATERNAL CIRCULATION

2005 ◽  
Vol 16 (2) ◽  
pp. 151-177 ◽  
Author(s):  
ALEC McEWAN
Keyword(s):  
Prenatal Diagnosis ◽  
Pregnant Women ◽  
Pulmonary Circulation ◽  
Blood Cells ◽  
White Blood Cells ◽  
Maternal Blood ◽  
Maternal Circulation ◽  
Fetal Cells ◽  
Y Chromosomes ◽  
Made In

Schmorl is generally credited with being the first to report finding fetal cells in maternal blood. In 1893 he identified trophoblasts in the pulmonary circulation of women who had died of eclampsia. Years later, in 1969, Walnowska identified Y chromosomes in lymphocytes isolated from the blood of pregnant women carrying male fetuses and this was repeated by Herzenberg in 1979 in white blood cells recognised as fetal by their surface HLA-A2 expression. Other sporadic reports followed but not until the 1990's did investigation into harvesting fetal cells from maternal blood begin in earnest. The aim of this article is to review the progress made in isolating and analysing these cells for the purposes of prenatal diagnosis.

scholarly journals A Monoclonal Antibody with Potential for Aiding Non-invasive Prenatal Diagnosis: Utility in Screening of Pregnant Women at Risk of Preeclampsia

2005 ◽  
Vol 53 (3) ◽  
pp. 345-350 ◽  
Author(s):  
Alejandra Fernández ◽  
Belén Prieto ◽  
Ana Escudero ◽  
Jack H. Ladenson ◽  
Francisco V. Alvarez
Keyword(s):  
Prenatal Diagnosis ◽  
Pregnant Women ◽  
Maternal Blood ◽  
Control Group ◽  
Maternal Circulation ◽  
Maternal Cell ◽  
Non Invasive ◽  
Nucleated Red Blood Cells ◽  
Increased Risk ◽  
Invasive Method

The development of a non-invasive method of prenatal diagnosis in maternal blood has been the goal of our investigations during the last years. We have developed several anti-CD71 monoclonal antibodies and optimized a protocol for the isolation of nucleated red blood cells (NRBC) from peripheral maternal blood. The enhanced traffic of fetal erythroblasts into the maternal circulation in preeclampsia has been investigated by several groups. In this study, we compared one of our antibodies, 2F6.3, with a commercial anti-CD71 antibody in blood samples from pregnant women suffering pregnancy-induced hypertension (PIH) and in a control group of pregnant women without clinical features suggestive of an increased risk of developing preeclampsia. The mAb 2F6.3, developed by our group, has succeeded in isolating a significantly higher number of erythroblasts with less maternal cell contamination than the commercial antibody in both women with PIH and in the control group ( p<0.01; Wilcoxon Signed Ranks Test). Florescence in situ hybridization analysis also demonstrated that 2F6.3 is a better antibody for the isolation of fetal NRBC in maternal blood than the commercial anti-CD71 antibody.

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