Purification of CD34~+/CD90~+ progenitor stem cells using immuno-magnetic microbeads selection and flow-cytometric sorting in peripheral blood leukapheresis

Objective To set up a practical method for purification of CD34 + stem cells and its subsets.Methods Aliquots of leukapheresis samples taken from 3 patients were firstly positive-selected for CD34 + stem cells by immuno-magnetic mircobeads,and then were purified for CD34 +/CD90 + progenitor stem cells using EPICS Elite fluorescence-activated cell sorter.Results The purity of selected CD34+ stem cells by immuno-magnetic separation was 83%-95% with the recovery rate of 54%-71% as well as the viability of more than 95%.After flow cytometric sorting,the purity and viability of CD34 +/CD90 + progenitor stem cell were 90% and 95% respectively,with a recovery of 40%-50%.The higher numbers of CD34 + stem cells in the beginning apheresis have,the higher purity and recovery of the procedures have.The morphologic characteristics between the two separations were different.Conclusions We established an effective procedure for purification of CD34 + stem cells and its subsets using combination of immuno-magnetic mircobeads selection and fluorescence-activated flow cytometric sorting,with satisfied cell purity,viability and recovery.