In vitro cultures of erythroid cells from maternal peripheral blood for the isolation of fetal cells

Several techniques for enrichment of nucleated fetal red blood cells presen t in maternal blood have been reported. Here we describe the use of in vitr o cultured erythroid cells from newborn cord blood and adult peripheral blo od. Together with a rapid high-performance liquid chromatography method tha t allows determination of as few as 100 cells containing hemoglobin (hb) F, quick optimization of enrichment methods for fetal nucleated red blood cel ls is possible. To generate pure erythroid progenitor cultures in vitro, th e nucleated cells from 20 mt of peripheral blood from pregnant women are gr own in media containing growth factors and hormones to yield hundreds of mi llions of immature erythroid cells within 2 weeks. In total, such cells can be passaged for more than 20 generations. These cells can either be mainta ined as BFU-E-like proerythroblasts or, by switching to media containing hu man recombinant erythropoetin plus insulin, be induced to mature into enucl eating erythrocytes, Thus, by varying the growth factor additions, their ma turation stages can be manipulated at will. Cells grown from adult peripher al blood have an increased amount of HbF (5-10%) compared to mature red cel ls from normal adult peripheral blood (HbF <1%), whereas cells expanded fro m newborn cord blood contain similar levels of HbF as cord blood erythrocyt es (70-80%).