Flow cytometric enumeration and immunophenotyping of hematopoietic stem and progenitor cells

Flow cytometric enumeration of CD34(+) hematopoietic stem and progenitor cells (HPC) is widely used to evaluate the adequacy of peripheral blood stem cell grafts and is also useful for planning the apheresis sessions needed to obtain these grafts. A state-of-the-art method to enumerate CD34(+) cells has been developed that makes use of a multiparameter definition of HPC, based on their light scatter characteristics and dim expression of CD45, utilizing fluorescent counting beads. This approach allows the absolute CD34(+) cell count to be determined directly from a flow cytometer. The method can be extended with a viability stain and additional markers for further immunologic characterization of CD34(+) cells, and has been successfully implemented in multicenter trials. Using such a standardized assay, it should be possible to define more accurately the lower threshold for a safe HPC graft in terms of short- and long-term hematopoietic reconstitution. Semin Hematol 38:139-147.