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.

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 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.

scholarly journals Enrichment, identification and analysis of fetal cells from maternal blood: evaluation of a prenatal diagnosis system

1999 ◽  
Vol 19 (7) ◽  
pp. 648-652 ◽  
Author(s):  
Irene M. de Graaf ◽  
Marja E. Jakobs ◽  
Nico J. Leschot ◽  
Ilya Ravkin ◽  
Simon Goldbard ◽  
...  
Keyword(s):  
Prenatal Diagnosis ◽  
Maternal Blood ◽  
Diagnosis System ◽  
Fetal Cells

scholarly journals In-Cell PCR Method for Specific Genotyping of Genomic DNA from One Individual in a Mixture of Cells from Two Individuals: A Model Study with Specific Relevance to Prenatal Diagnosis Based on Fetal Cells in Maternal Blood

2002 ◽  
Vol 48 (12) ◽  
pp. 2115-2123 ◽  
Author(s):  
T Vauvert Hviid
Keyword(s):  
Prenatal Diagnosis ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Chromosomal Abnormalities ◽  
Maternal Blood ◽  
Specific Sequence ◽  
Mixed Cell ◽  
Fetal Cells ◽  
Model Study ◽  
Conventional Cell

Abstract Background: During recent years, much attention has been paid to the possibility of using fetal cells circulating in the pregnant woman’s blood for prenatal diagnosis of genetic or chromosomal abnormalities. Although successes have been achieved in enrichment procedures for fetal cells from maternal blood samples, the use of such an approach for genotyping by molecular biology techniques in a more routine setting has been hampered by the large contamination of maternal nucleated blood cells in the cell isolates. Therefore, a new method based on in-cell PCR is described, which may overcome this problem. Methods and Results: Mixtures of cells from two different individuals were fixed and permeabilized in suspension. After coamplification of a DNA sequence specific for one of the individuals and the DNA sequence to be genotyped, the two PCR products were linked together in the fixed cells positive for both DNA sequences by complementary primer tails and further amplification steps. In a model system of mixtures of male and female CD71-positive cells from umbilical cord blood attached to immunomagnetic particles, a Y-chromosome-specific sequence (TSPY) was linked to a polymorphic HLA-DPB1 sequence only in the male cells, leading to the correct HLA-DPB1 genotyping of the male by DNA sequencing of a nested, linked TSPY-HLA-DPB1 PCR product. Conclusion: This approach might be usable on mixed cell populations of fetal and maternal cells obtained after conventional cell-sorting techniques on maternal peripheral vein blood.

Sign in / Sign up

Export Citation Format

Share Document