Comparison of fetal cell recovery from maternal blood using a high density gradient for the initial separation step: 1.090 versus 1.119 g/ml
✍ Scribed by Osamu Samura; Akihiko Sekizawa; Dong Kai Zhen; Vincent M. Falco; Diana W. Bianchi
- Publisher
- John Wiley and Sons
- Year
- 2000
- Tongue
- English
- Weight
- 67 KB
- Volume
- 20
- Category
- Article
- ISSN
- 0197-3851
No coin nor oath required. For personal study only.
✦ Synopsis
The purpose of this study was to improve recovery of fetal nucleated erythrocytes (NRBCs) from maternal blood for non-invasive prenatal diagnosis. Peripheral blood samples were obtained from 27 women who had just undergone pregnancy termination at 6 to 23 weeks. Samples were split and mononuclear cells were isolated using Histopaque gradient at densities of 1.090 g/ml and 1.119 g/ml. CD45 depletion using magnetic activated cell-sorting, followed by ¯ow-sorting with antibody to c-globin and ¯uorescence in situ hybridization (FISH) analysis, were used to evaluate the number of fetal NRBCs recovered. In samples separated with the 1.119 g/ml density gradient, the yield of true anti-c haemoglobin positive cells (median, 14.9; range, 0±717.5) was signi®cantly higher than that with the 1.090 g/ml density gradient (median, 4.9; range, 0±532.5). After FISH analysis, in the 14 samples in which the fetal karyotype differed from the mother, the median number of fetal NRBCs separated by the 1.119 g/ml density gradient was 22.9 (2±717.5), which was signi®cantly higher than that by the 1.090 g/ml gradient (median, 11.5; range, 0±532.5, p=0.022). Increased density of the gradient used for the initial enrichment of fetal cells results in improved fetal cell recovery in fresh post-termination blood samples, which may permit better non-invasive detection of fetal cells in maternal blood.