Flow cytometric enumeration and immunophenotyping of hematopoietic stem and progenitor cells

Abstract

Flow cytometric enumeration of CD34+ hematopoietic stem and progenitor cells (HPC) is widely used for evaluation of graft adequacy of peripheral blood stem cell grafts, and is also useful in planning the apheresis sessions necessary to obtain these grafts. The state-of-the-art method to enumerate CD34+ cells makes use of a multiparameter definition of HPC based on their light scatter characteristics and dim expression of CD45, and the use of counting beads to derive the concentration of CD34+ cells directly from the flow cytometric assessment. This method can be extended with a viability stain and additional markers for further immunological characterization of CD34+ cells, and has been successfully implemented in multicenter trials. Thus, the lower threshold of a safe HPC graft in terms of short- and long-term hematopoiesis may be more accurately defined.