A comparison of different density gradients and antibodies for enrichment of fetal erythroblasts by MACS.
The enrichment of fetal cells, in particular fetal erythroblasts from the blood of pregnant women offers a promising non-invasive alternative for prenatal diagnosis. The purpose of this study was to compare the retrieval of erythroblasts by different density gradients and different antibodies against erythroid surface antigens, in both a model test system and in blood samples of pregnant women. We enriched erythroblasts from artificial mixtures of cord and adult blood (1:50) and from 16 ml of peripheral blood from pregnant women at a mean gestational age of 14+2 weeks. The yield of erythroblasts was calculated and compared using Wilcoxon's matched-pairs signed-rank test. In the artificial mixture most erythroblasts were retrieved using the heaviest density gradient (specific density of 1119) followed by antibody-labelled magnetic cell sorting (MACS). With anti-GPA the yield of erythroblasts from the artificial mixture was highest (9362.5 erythroblasts) compared with anti-CD36 (5164.3), anti-i (3455.2), anti-CD71 (3055.8) and HAE9 (2364.2). The difference between anti-GPA and anti-CD71, HAE9 and anti-i was significant (p=0.0277). The enrichment of erythroblasts from peripheral blood of pregnant women showed similar results. The yield of erythroblasts using anti-GPA was the highest. These results enable us to simplify our enrichment protocols to a single density gradient of 1119 specific density followed by MACS, with anti-GPA.
Department of Obstetrics and Gynaecology, University of Basel, Switzerland.
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Troeger
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Holzgreve
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Hahn
D000911:Antibodies, Monoclonal / Q000276:immunology*; D015703:Antigens, CD / Q000276:immunology; D000944:Antigens, Differentiation, B-Lymphocyte / Q000276:immunology; D000954:Antigens, Surface / Q000276:immunology; D018955:CD36 Antigens / Q000276:immunology; D002499:Centrifugation, Density Gradient; D004900:Erythroblasts / Q000276:immunology*; D005260:Female; D005312:Fetal Blood / Q000166:cytology; D005333:Fetus / Q000166:cytology*; D005865:Gestational Age; D006021:Glycophorins / Q000276:immunology; D006801:Humans; D007048:I Blood-Group System / Q000276:immunology; D018189:Immunomagnetic Separation / Q000379:methods*; D011247:Pregnancy; D011990:Receptors, Transferrin; D018709:Statistics, Nonparametric
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; D000954:Antigens, Surface; D018955:CD36 Antigens; C068126:CD71 antigen; D006021:Glycophorins; D007048:I Blood-Group System; D011990:Receptors, Transferrin