A comparison of different density gradients and antibodies for enrichment of fetal erythroblasts by MACS

Prenat Diagn. 1999 Jun;19(6):521-6.

Abstract

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antibodies, Monoclonal / immunology*
  • Antigens, CD / immunology
  • Antigens, Differentiation, B-Lymphocyte / immunology
  • Antigens, Surface / immunology
  • CD36 Antigens / immunology
  • Centrifugation, Density Gradient
  • Erythroblasts / immunology*
  • Female
  • Fetal Blood / cytology
  • Fetus / cytology*
  • Gestational Age
  • Glycophorins / immunology
  • Humans
  • I Blood-Group System / immunology
  • Immunomagnetic Separation / methods*
  • Pregnancy
  • Receptors, Transferrin
  • Statistics, Nonparametric

Substances

  • Antibodies, Monoclonal
  • Antigens, CD
  • Antigens, Differentiation, B-Lymphocyte
  • Antigens, Surface
  • CD36 Antigens
  • CD71 antigen
  • Glycophorins
  • I Blood-Group System
  • Receptors, Transferrin