PHENOTYPING AND SORTING OF MURINE BONE MARROW HAEMATOPOIETIC STEM CELLS USING FLOW CYTOMETRY

To develop a protocol of multiparametric phenotyping and sorting of LSK-subpopulations of hematopoietic stem cells and to determine their relative numbers in the bone marrow of mice was the goal of this research. The modified protocol of multiparametric phenotyping of murine hematopoietic stem cells enable to determine the content of Lin?Sca-1+ c-kit+, Lin?Sca-1+c-kit+flt3+CD150?, Lin?Sca-1+c-kit+flt3+CD150+ and Lin?Sca-1+c-kit+flt3?CD150? subpopulations in bone marrow of FVB mice. It was shown that the dominant population among LSK-cells represents the phenotype Lin?Sca-1+c-kit+flt3?CD150? (57.2 ± 6.8%), which characterizes the short-term hematopoietic stem cells responsible for myelopoiesis. Also the protocol of sorting of murine bone marrow LSK-cells was proposed and its effectiveness for subsequent transplantation in experiments was demonstrated. At repeated phenotyping of sorted cells the purity of Lin?Sca-1+c-kit+ cell fraction was 96.6 ± 1.8% with viability up to 89.6 ± 4.6%.