Enrichment of fetal nucleated red blood cells from maternal peripheral blood by using density gradient centrifugation and magnetically activated cell sorting

Objective To sort and enrich fetal nucleated red blood cells(NRBCs)from maternal peripheral blood by using discontinuous density gradient centrifugation(DGC) and magnetically activated cell sorting(MACS).Methods A total of 21 pregnant women with a gestation of 12 to 28 weeks were studied in this paper,10 ml peripheral blood was obtained from each pregnant woman,NRBCs were sorted by methods of DGC and MACS.Meanwhile,smear slides of maternal peripheral blood before and after enrichment step were subjected to Wright's staining and the staining of antibody against the fetal hemoglobin(HbF).Results No NRBCs was found in the Wright's staining slides of maternal blood without enriching NRBCs.Enrichment of NRBCs by DGC method,there were 1～8 NRBCs which were found in 11 of 21 maternal samples;Combined DGC method with MACS,there were 1～7 NRBCs in 12 of 21 maternal samples respectively.And the staining results using antibody against HbF confirmed that fetal NRBCs exist in the slides.Conclusion Our study had primarily established the efficient method of sorting and enrichment of fetal NRBCs from maternal blood,it can be applied in the further prenatal genetic diagnosis and other research work.