Isolation of fetal erythroid cells from maternal blood based on expression of erythropoietin receptors.
Fetal nucleated red cells which pass into the maternal circulation during pregnancy are a potential cell source for non-invasive prenatal genetic diagnosis. To sort these rare cells with a high degree of specificity, we focussed our attention on the erythropoietin receptor, a strictly erythroid-specific antigen. We first labelled these receptors with biotin-(sialyl)-erythropoietin, then isolated the erythroid cells by magnetic beads conjugated with streptavidin in a MiniMACS (magnetic cell separator). The effectiveness of this strategy for the enrichment of fetal cells was evaluated by assessing its accuracy for gender prediction in 18 male-bearing pregnancies. Polymerase chain reaction (PCR) results on maternal blood samples sorted for Epo-r and CD71 antigens displayed similar sensitivity (55% Epo-r, 61% CD71) in detecting Y-specific sequences while immunocytochemical studies on four maternal blood samples, sorted after increasing the binding time of the ligand to Epo-r (8 h), showed a substantial improvement in fetal cell recovery and purity. We conclude that sorting by Epo-r/biotin-(sialyl)-erythropoietin provides effective enrichment of fetal nucleated red cells allowing the possibility of direct prenatal cytogenetic analysis by multiprobe fluorescent in-situ hybridization (FISH).
Valerio
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Istituto di Ricerche Genetiche, V.le della Costituzione, Napoli, Italy.
Aiello
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Altieri
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D015703:Antigens, CD / Q000235:genetics; D000944:Antigens, Differentiation, B-Lymphocyte / Q000235:genetics; D002469:Cell Separation / Q000379:methods; D004912:Erythrocytes / Q000166:cytology* / Q000378:metabolism*; D005260:Female; D005312:Fetal Blood / Q000166:cytology* / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D008431:Maternal-Fetal Exchange; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011296:Prenatal Diagnosis; D017467:Receptors, Erythropoietin / Q000235:genetics / Q000378:metabolism*; D011990:Receptors, Transferrin; D012680:Sensitivity and Specificity; D012732:Sex Determination Analysis / Q000379:methods; D014998:Y Chromosome / Q000235:genetics
D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; C068126:CD71 antigen; D017467:Receptors, Erythropoietin; D011990:Receptors, Transferrin