ABSTRACT
Objective The effectiveness on the enrichment of nuclear red blood cells(NRBC) in umbilical cord blood by polyvinylidene difluoride membrane was investigated with intervention on membrane ionic channels.Methods NRBC was isolated by the isolation solution with a density of 1.067 g/ml.The rates of the cell enrichment using verapamil and furosemide intervention by discontinuous density centrifugation were examined by flow cytometry.The NRBC that past through PVDF membrane with pore size of 5 ?m were checked and quantified by nested-PCR.Results The optimal centrifugation density was 1.067 g/ml.The mean purity of NRBC after centrifugation was 2.54%,yet the purity after intervention increased to 9.36%(3.7-fold increment).The NRBC passing through PVDF membrane was from 0.83% to 6.15%,with enrichment of 7.4 times.When NRBC was 120 cells/ml,the density of the nested PCR products increased by nearly 4 times after intervention(4.48 vs 17.78).Conclusions Verapamil and furosemide could affect the morphology of cells and promote the rate of NRBC enrichment.