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Fetal Cells from Maternal Blood - A Powerful tool for Prenatal Diagnosis
(Abstract published in the proceedings of ISPD 2010, Amsterdam)
Marie Brinch, Lotte Hatt, Ripudaman Singh, Kristine Møller, Rune Hoff Lauridsen, Helle Kristensen, Maja B. Kristensen, Marianne Mose Hansen, Marianne Rasmussen, Simon Tabi Arrey, Trine Overgaard Petersen, Steen Kølvraa, Britta Christensen*
FCMB ApS. Tysklandsvej 7. Vejle 7100. Denmark. Email:
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*
Circulating fetal cells in maternal blood provide a powerful tool for risk-free, non-invasive prenatal diagnosis of fetal anomalies. Since late 19th century, scientists have known that fetal cells do circulate in maternal blood. However, due to lack of knowledge on fetal cell specific markers and suitable methods to enrich these rare cells, the potential of circulating fetal cells for non-invasive prenatal diagnosis has not been fully acknowledged. We at FCMB have developed a novel method to enrich rare fetal cells from maternal blood. The fetal origin of the enriched cells was confirmed by doing XY chromosome-specific FISH on pregnancies bearing a male fetus. We used 100 fetal cells (in a biological replicate of 2) to prepare cDNA for microarray analysis. A comparison of the array data from the fetal cells and the surrounding maternal blood cells revealed unique fetal cell markers which were used for gender-independent isolation/identification of fetal cells from maternal blood. To confirm this we performed a feasibility study where intravenous blood was collected from 100 pregnant women (gestation age 11 to 14 weeks). Fetal cells were isolated from maternal blood by magnetic cell sorting using ‘FCMB cocktail-1’ antibodies. The cells were stained using ‘FCMB cocktail-2’ and later identified by automated scanning. The gender of the fetal cells identified by antibody staining was determined by XY-FISH and matched with gender determined by Y specific PCR on free fetal DNA from maternal plasma. There was a 99% correlation between the results of gender analysis on fetal cells by XY-FISH and gender analysis on free fetal DNA, confirming our ability to enrich and detect fetal cells in a gender-independent manner. Taking lead from our ability to successfully isolate fetal cells, we are now doing a feasibility study on high risk pregnancies. |
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Isolation of Fetal Cells from Maternal Blood - From Fact to Reality
(Abstract presented at IFPA 2009, Adelaide)
Ripudaman Singh*, Andreas Eckelt, Ann Birte Bro, Britta Christensen, Hanne Skannerup, Helle Kristensen, Kristine Møller, Lotte Hatt, Maja B. Kristensen, Marianne Rasmussen, Marie Brinch, Otto Bjerg Hausgaard, Rune Hoff Lauridsen, Simon Tabi Arrey, Steen Kølvraa, Trine Overgaard Petersen.
FCMB ApS. Tysklandsvej 7. Vejle 7100. Denmark. *Email:
This e-mail address is being protected from spambots. You need JavaScript enabled to view it
“Foreign cells from the fetus do circulate in mother’s blood” – This was the fact known to the biologists since late 19th century when Georg Schmorl first described the presence of trophoblast sprouts in the lungs of women who died of eclampsia. It has taken biologists another century to decipher methods to isolate the rare fetal cells from maternal circulation, which have great potential in non-invasive prenatal diagnosis. We have developed a novel method of consistently separating rare fetal cells from maternal circulation. Intravenous blood was collected from pregnant women (gestation age 11 to 14 weeks). Gender of the fetus was checked by performing Y specific PCR. 24ml blood from mothers carrying male fetus was prepared for antibody labeling using a proprietary preparation technology. The cells were then incubated with ‘FCMB cocktail-1’ antibodies, fetal cells isolated by magnetic cell sorting and later identified by automated scanning of XY-FISH. 754 fetal cells, on an average of 1cell/5ml of whole blood, were isolated. Some cells were rehybridized using reverse-colour XY-FISH to confirm their fetal origin. 100 fetal cells (in a biological replicate of 2) were used to prepare cDNA for microarray analysis. A comparison of the array data from the fetal cells with the control maternal blood cells gave us a clear indication of the unique markers that are expressed in the fetal cells. We have based this knowledge to further develop a method for isolation/detection of fetal cells from maternal circulation. We have also been successful in staining fetal cells with a cocktail of intracellular antibodies. This has further enhanced our ability to perform a gender-free isolation/detection of fetal cells from maternal blood. Taking a lead from our ability to successfully and consistently isolate fetal cells, we are now in the process of doing a feasibility study on high risk pregnancies. |
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FCMB Publications 
