SELECTION OF FETAL CELLS CIRCULATING IN MATERNAL BLOOD FOR NONINVASIVE PRENATAL DIAGNOSIS | Выделение клеток плода, циркулирующих в крови матери, для неинвазивной пренатальной диагностики
2016
Gudkov G. V. | Filippov E. F. | Krutenko D. V.
Trophoblast cells circulating in maternal blood may serve as potential sources of genetic information for screening in obstetric practice approaches to noninvasive prenatal diagnosis. The material for the study was whole blood of pregnant women of gestational ages of 8-12 weeks in a volume of 10 ml. stabilized with heparin. Enrichment was performed by the method of gradient centrifugation and magnetic separation (CD45–) with fluorescent staining negative fractions labeled with antibodies to antigens of trophoblasts (HLA-G, and Trop-2) to conduct flow cytometry and sorting of cells on glass slides. Single cell candidates using laser micro dissection were transferred into a vial for subsequent whole-genome amplification, providing sufficient representativeness of their genome. Fetal origin of the genetic material was confirmed by comparison of alleles of the HLA genes of the parents and cells candidates. It is possible to improve significantly the accuracy and versatility of non-invasive prenatal diagnosis using comparative genomic hybridization using chips (аCGH)
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