Enrichment of Fetal Nucleated Red Blood Cells from the Maternal Circulation for Prenatal Diagnosis: Experiences with Triple Density Gradient and MACS Based on More than 600 Cases

Objective: We wanted to obtain statistically relevant data about the efficiency of our method for the isolation of fetal nucleated red blood cells (NRBCs) from the maternal circulation. Methods: More than 600 samples were investigated using a triple density gradient followed by magnetic separation of anti-CD71-labeled cells, and yields and purities of recovered NRBCs were determined. Results: The enrichment effectivity as well as the morphological condition of cells was reproducibly good, if blood samples were enriched within 48 h after sampling. The efficacy was independent of various methodological parameters and our technique was superior to other magnetic cell-sorting techniques. Mean yields and purities of NRBCs increased with increasing gestational age, ranging from 100 to 1,000 cells per 40-ml blood sample and from 0.1 to 1%, respectively, from the 6th week of gestation to term. In pregnancies with preeclampsia NRBCs were increased by a factor of 10. Conclusion: Our enrichment technique proved to be optimized with respect to various methodological parameters, which were compared in the present study, and it is efficient and reproducible for the enrichment of NRBCs from the maternal circulation in all three gestational trimesters.

[1]  D. Rubinsztein,et al.  Enrichment of fetal nucleated cells from maternal blood: Model test system using cord blood , 1995, Prenatal diagnosis.

[2]  D. Bianchi.  Clinical Trials and Experience: Boston , 1994, Annals of the New York Academy of Sciences.

[3]  G. Weil,et al.  Male fetal progenitor cells persist in maternal blood for as long as 27 years postpartum. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[4]  S. Wachtel,et al.  Fetal cells in maternal blood: recovery by charge flow separation , 1996, Human Genetics.

[5]  I. Sargent,et al.  Presence of fetal DNA in maternal plasma and serum , 1997, The Lancet.

[6]  S. Latt,et al.  Analysis of peripheral maternal blood samples for the presence of placenta‐derived cells using Y‐specific probes and McAb H315 , 1988, Prenatal diagnosis.

[7]  S. Latt,et al.  Isolation of fetal DNA from nucleated erythrocytes in maternal blood. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[8]  W. Holzgreve,et al.  Prenatal diagnosis using fetal cells in the maternal circulation , 1995 .

[9]  W. Holzgreve,et al.  Detection of Fetal Trisomies 21 and 18 From Maternal Blood Using Triple Gradient and Magnetic Cell Sorting , 1993, American journal of reproductive immunology.

[10]  A. Morley,et al.  Isolation of fetal trophoblast cells from peripheral blood of pregnant women , 1990, The Lancet.

[11]  C. Rodeck,et al.  The time of appearance, and quantitation, of fetal DNA in the maternal circulation. , 1994, Annals of the New York Academy of Sciences.

[12]  H. Takabayashi,et al.  Development of non‐invasive fetal DNA diagnosis from maternal blood , 1995, Prenatal diagnosis.

[13]  S. Elias,et al.  Prenatal Diagnosis of Aneuploidy Using Fetal Cells Isolated from Maternal Blood University of Tennessee, Memphis Experience a , 1994, Annals of the New York Academy of Sciences.

[14]  K. Benirschke,et al.  Anatomical Relationship between Fetus and Mother , 1994, Annals of the New York Academy of Sciences.

[15]  K. Klinger,et al.  Prenatal diagnosis by use of fetal cells isolated from maternal blood. , 1995, American journal of obstetrics and gynecology.

[16]  W. Holzgreve,et al.  Magnetic cell sorting and the transferrin receptor as potential means of prenatal diagnosis from maternal blood. , 1992, American journal of obstetrics and gynecology.

[17]  A. Covone,et al.  TROPHOBLAST CELLS IN PERIPHERAL BLOOD FROM PREGNANT WOMEN , 1984, The Lancet.

[18]  S Miltenyi,et al.  Enrichment of fetal cells from maternal blood by high gradient magnetic cell sorting (double MACS) for PCR‐based genetic analysis , 1994, Prenatal diagnosis.

[19]  W. Holzgreve,et al.  Detection of fetal DNA in maternal blood by PCR , 1990, The Lancet.

[20]  W. Holzgreve,et al.  Fetal DNA in uterine vein blood , 1992, Obstetrics and gynecology.

[21]  Leonore A. Herzenberg,et al.  Fetal cells in the blood of pregnant women: detection and enrichment by fluorescence-activated cell sorting. , 1979, Proceedings of the National Academy of Sciences of the United States of America.