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For simple and effective isolation of fetal cells from peripheral maternal blood, we combined depletion of maternal cells and enrichment of fetal cells by high-gradient magnetic cell separation (MACS). First CD45+ and CD14+ cells were depleted from maternal peripheral blood mononuclear cells by MACS. From the depleted fraction, CD71+ erythroid cells were enriched up to 80 per cent by MACS. This double-MACS' procedure yielded an average depletion rate of 780-fold and an average enrichment rate of 500-fold, with approximate recovery rates of 40-55 per cent. For paternity testing, cells from unseparated blood and the various fractions were analysed for polymorphism of the HLA-DQ-A1 locus and D1S80 locus by the polymerase chain reaction (PCR). In CD45-/CD71+ sorted cells from maternal blood, but not in unfractionated cells from maternal blood or CD45-/CD14- cells, paternal alleles could be detected. In the CD45-/CD71+ fraction, the relative frequency of paternal alleles compared with maternal alleles ranged from 1 in 20 to 1 in 200 (determined by titration and depending on the quality of separation and biological variation). In 7 out of 11 cases, between weeks 12 and 25 of gestation, we could identify paternal alleles by PCR, either HLA-DQ-A1 or D1S80. This double-MACS procedure is simple, fast, efficient, and reliable for non-invasive prenatal diagnosis.
J Büsch, and
P Huber, and
E Pflüger, and
S Miltenyi, and
J Holtz, and
A Radbruch
June 2002,
Congenital anomalies,
J Büsch, and
P Huber, and
E Pflüger, and
S Miltenyi, and
J Holtz, and
A Radbruch
April 2000,
Archives of gynecology and obstetrics,
J Büsch, and
P Huber, and
E Pflüger, and
S Miltenyi, and
J Holtz, and
A Radbruch
January 1990,
Cytometry,
J Büsch, and
P Huber, and
E Pflüger, and
S Miltenyi, and
J Holtz, and
A Radbruch
September 2002,
Congenital anomalies,
J Büsch, and
P Huber, and
E Pflüger, and
S Miltenyi, and
J Holtz, and
A Radbruch
January 1993,
American journal of reproductive immunology (New York, N.Y. : 1989),
J Büsch, and
P Huber, and
E Pflüger, and
S Miltenyi, and
J Holtz, and
A Radbruch
January 1998,
Fetal diagnosis and therapy,
J Büsch, and
P Huber, and
E Pflüger, and
S Miltenyi, and
J Holtz, and
A Radbruch
June 2001,
The journal of obstetrics and gynaecology research,
J Büsch, and
P Huber, and
E Pflüger, and
S Miltenyi, and
J Holtz, and
A Radbruch
January 1994,
Methods in cell biology,
J Büsch, and
P Huber, and
E Pflüger, and
S Miltenyi, and
J Holtz, and
A Radbruch
January 1998,
In vivo (Athens, Greece),
J Büsch, and
P Huber, and
E Pflüger, and
S Miltenyi, and
J Holtz, and
A Radbruch
January 2000,
Methods in molecular biology (Clifton, N.J.),
