Fetal cells in maternal blood: Determination of purity and yield by quantitative polymerase chain reaction

Bianchi, Diana W.; Shuber, Anthony P.; DeMaria, Mary Ann; Fougner, Arthur C.; Klinger, Katherine W.

doi:10.1016/s0002-9378(94)70059-1

Cited by 80 publications

(41 citation statements)

References 17 publications

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“…The fact that the persisting cells were detected only by Southern hybridization or nested PCR techniques implies that they are extremely rare and probably circulate in maternal blood at much lower frequencies than Schroder et al (4) originally suggested. We have independently described techniques of fetal cells quantitation by PCR that might be applied to future experiments to address the question of frequency (13).…”

Section: Discussionmentioning

confidence: 99%

Male fetal progenitor cells persist in maternal blood for as long as 27 years postpartum.

Bianchi

Zickwolf

Weil

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

1,154

788

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Rare nucleated fetal cells circulate within maternal blood. Noninvasive prenatal diagnosis by isolation and genetic analysis of these cells is currently being undertaken. We sought to determine if genetic evidence existed for persistent circulation of fetal cells from prior pregnancies. Venous blood samples were obtained from 32 pregnant women and 8 nonpregnant women who had given birth to males 6 months to 27 years earlier. Mononuclear cells were sorted by flow cytometry using antibodies to CD antigens 3, 4, 5, 19, 23, 34, and 38. DNA within sorted cells, amplified by PCR for Y chromosome sequences, was considered predictive of a male fetus or evidence of persistent male fetal cells. In the 32 pregnancies, male DNA was detected in 13 of 19 women carrying a male fetus. In 4 of 13 pregnancies with female fetuses, male DNA was also detected. All of the 4 women had prior pregnancies; 2 of the 4 had prior males and the other 2 had terminations of pregnancy. pearance. In a group of 20 primigravidas sampled on more than one occasion postpartum, quinacrine fluorescent signals ("Y body") were seen at a frequency of 0.01-0.1% of lymphocytes. These frequencies were noted to remain "practically unchanged" up until 1 year after delivery, when the study ended. In a related study, Ciaranfi et al. (5) (4) to include samples from women 5-7 years postpartum (5). In a series of 62 women who delivered a male infant, more than half had detectable male lymphocytes 2 years after birth. In some cases, metaphases with a Y chromosome were visualized 5 years postpartum. These studies, although intriguing, were performed in the 1970s using techniques less sensitive and less accurate than those available today.During this decade, investigators using PCR amplification of Y chromosome-specific sequences to identify fetal cells in maternal blood noted "false positive" results-i.e., detection of male DNA when the fetus was female (6, 7). Although laboratory contamination as a source of true false positives can be a concern (8), it appears that, in some cases, the male DNA was a real finding, possibly originating from a prior pregnancy (9). Other investigators studied postpartum maternal samples without using cell separation techniques to enrich for specific subpopulations of fetal cells and concluded that fetal cells did not persist postpartum (10).The study reported here occurred in two parts. Initially, we used a monoclonal antibody to cluster differentiation antigen (CD) 34 to isolate fetal hematopoietic stem cells from the blood of pregnant women. The surprising results, described below, suggested that the gender of the circulating CD34+ cells did not always correlate with the fetal gender of the current pregnancy. This led to the hypothesis that fetal CD34+ cells could persist from a prior pregnancy. We tested this hypothesis by fluorescence-activated cell sorting of four specific subpopulations of mononuclear cells from women who were not presently pregnant but who had previously given birth to a male infant. Although we were in...

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Section: Discussionmentioning

confidence: 99%

Male fetal progenitor cells persist in maternal blood for as long as 27 years postpartum.

Bianchi

Zickwolf

Weil

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

1,154

788

View full text Add to dashboard Cite

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“…Bianchi et al [11][12][13][14][15] used quantitative PCR to detect male fetal DNA from samples following fluorescence-activated sorting with various combinations of monoclonal antibodies. Even though the PCR approach is highly sensitive, it amplifies all male-specific DNA.…”

Section: Discussionmentioning

confidence: 99%

Frequencies of Fetal Nucleated Red Blood Cells in Maternal Blood during Different Stages of Gestation

Kuo

1998

Fetal Diagn Ther

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Nucleated red blood cells (NRBCs) isolated by a triple density gradient from 100 ml peripheral blood samples of 100 pregnant women and 30 women postpartum were subjected to morphological analysis and PCR quantitation. The number of NRBCs steadily increased from 5.3 (frequency: 2.4 × 10^–7) in early gestation to 98.2 (frequency: 4.2 × 10^–6) near term. The number of male cells increased from 6 (frequency: 2.7 × 10^–7) in early gestation to a peak of 31 (frequency: 1.48 × 10^–6) in the second trimester, and slightly decreased to 27 (frequency: 1.31 × 10^–6) near term. Both NRBCs and male fetal cells rapidly disappeared after delivery. The result implies that a significant proportion of NRBCs in maternal blood are of fetal origin before 24 weeks of gestation while in late gestation the majority of NRBCs may be of maternal origin.

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“…However, it is not fully known if fetal cells are present in sufficient number in all pregnant women. The estimated number of fetal nucleated red blood cells circulating in maternal blood is very low (Bianchi et al, 1994;Reading et al, 1995;Takabayashi et al, 1995;Bianchi et al, 1997). In order to analyse these cells one needs to enrich them using one of several methods previously described in the literature (Hahn et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

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

et al. 1999

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Fetal cells in maternal blood: Determination of purity and yield by quantitative polymerase chain reaction

Cited by 80 publications

References 17 publications

Male fetal progenitor cells persist in maternal blood for as long as 27 years postpartum.

Male fetal progenitor cells persist in maternal blood for as long as 27 years postpartum.

Frequencies of Fetal Nucleated Red Blood Cells in Maternal Blood during Different Stages of Gestation

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

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