Enrichment of fetal trophoblasts and nucleated erythrocytes from maternal blood by an immunomagnetic colloid system.
The aim of this study was to isolate fetal trophoblasts and nucleated erythrocytes from maternal blood using the immunomagnetic colloid system. About 25 ml of maternal blood was collected from pregnant women between of 14 and 20 weeks gestation. Nucleated erythrocytes (NRBCs) were isolated from 5 ml of maternal blood and a nested polymerase chain reaction for the Y chromosome was used to determine fetal origin. The sensitivity of the fetal gender diagnosis was 80% and the specificity was 86%. Both fetal trophoblasts and NRBCs were isolated from the remaining 20 ml of maternal blood. The fetal gender of the trophoblast-enriched fraction was determined using fluorescence in situ hybridisation (FISH) with dual-colour XY-specific DNA probes. XY-specific signals were observed in 0.38% of cells sorted from all pregnant women carrying male fetuses (n = 10). Simultaneous immunophenotyping for the fetal haemoglobin and FISH using XY probes were used to evaluate the fetal origin of cells enriched with anti-CD71. The mean percentage of male fetal erythroblasts was 0.24% and the number of fetal erythroblasts was estimated to be about 672 in 20 ml of maternal blood. The number of fetal erythroblasts detected in our study was greater than that detected by most other separation techniques. Our study shows that it would be feasible to use the immunomagnetic colloid system for the isolation of both trophoblasts and NRBCs from the same maternal blood sample with relatively good efficiency.
Department of Obstetrics and Gynaecology, Singapore General Hospital, Singapore. gogtyc@sgh.gov.sg
T H
TH
Lim
A
A
Tan
V H
VH
Goh
D000649:Amniocentesis / Q000379:methods; D015703:Antigens, CD / Q000276:immunology; D000944:Antigens, Differentiation, B-Lymphocyte / Q000276:immunology; D003102:Colloids; D015342:DNA Probes; D004912:Erythrocytes / Q000166:cytology*; D005260:Female; D005319:Fetal Hemoglobin / Q000032:analysis*; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods; D016130:Immunophenotyping; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D008297:Male; D011247:Pregnancy; D011990:Receptors, Transferrin; D012680:Sensitivity and Specificity; D012732:Sex Determination Analysis / Q000379:methods*; D014327:Trophoblasts / Q000166:cytology*; D014998:Y Chromosome
D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; C068126:CD71 antigen; D003102:Colloids; D015342:DNA Probes; D011990:Receptors, Transferrin; D005319:Fetal Hemoglobin