New method for stem cell quantification: applications to the management of peripheral blood stem cell transplantation

Abstract

A dramatic increase in peripheral blood stem cells (PBSC) is observed after high-dose chemotherapy followed by haematopoietic growth factors. The degree of mobilisation of PBSC is quantified by the level of clonogenic cells detected by CFU assays (CFU-GM or CFU-GEMM) or CD34+ cell determination. Working under the hypothesis that, in peripheral blood, mononuclear cells in DNA synthesis (MCDS) are proliferating stem cells, we decided to detect these cells by flow cytometric measurement of their DNA content. The relations between the number of MCDS and well-known haematopoietic progenitor indicators such as CFU-GM or CD34+ cells were analysed. We studied the kinetics of recruitment of PBSC in cancer patients, treated with rmeHuG-CSF following VP-16 cytoxan chemotherapy, until the first day of leukapheresis. For the 31 patients studied the individual curves of peripheral MCDS and CFU-GM reconstitutions showed identical profiles and a good correlation was noted between the numbers of peripheral MCDS and CFU-GM (r = 0.73). In the leukapheresis product, the predictive value of MCDS was equivalent to CFU-GM for PBSC quantification (r = 0.70). In conclusion, MCDS analysis by flow cytometry provides reliable results and appears to be an alternative to CFU-GM assay or CD34+ cell determination for PBSC quantification.