Enrichment of fetal trophoblasts and nucleated erythrocytes from maternal blood by an immunomagnetic colloid system

Citation
Th. Lim et al., Enrichment of fetal trophoblasts and nucleated erythrocytes from maternal blood by an immunomagnetic colloid system, HUM GENET, 104(5), 1999, pp. 399-404
Citations number
20
Categorie Soggetti
Molecular Biology & Genetics
Journal title
HUMAN GENETICS
ISSN journal
03406717 → ACNP
Volume
104
Issue
5
Year of publication
1999
Pages
399 - 404
Database
ISI
SICI code
0340-6717(199905)104:5<399:EOFTAN>2.0.ZU;2-4
Abstract
The aim of this study was to isolate fetal trophoblasts and nucleated eryth rocytes 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 chromos ome 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 mi of maternal blood. The fetal gender of the trophoblast-enriched fraction was determined using fluorescen ce 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 th e fetal haemoglobin and FISH using XY probes were used to evaluate the feta l origin of cells enriched with anti-CD71. The mean percentage of male feta l erythroblasts was 0.24% and the number of fetal erythroblasts was estimat ed to be about 672 in 20 ml of maternal blood. The number of fetal erythrob lasts detected in our study was greater than that detected by most other se paration techniques. Our study shows that it would be feasible to use the i mmunomagnetic colloid system for the isolation of both trophoblasts and NRB Cs from the same maternal blood sample with relatively good efficiency.