Abstract

The hematopoiesis steadily generates a large number of blood cells by intensive cell proliferation. A hierarchy of hematopoietic stem and progenitor cells (HSPCs) has been well characterized in mouse. The aim of our study was to find out to which extent these distinct populations contributed to blood cell production. We estimated the production rates of various types of HSPCs according to the percentage of S phase cells, their number in bone marrow, and the S-phase duration. Flow cytometry was used for immunophenotyping of HSPCs according to Lin markers, c-Kit, Sca-1, CD48, CD150, CD34, CD16/32, CD71, and IL7R expression. The dual BrdU/EdU labelling technique was applied to determine the S-phase duration and the percentage of DNA-synthesizing (S-phase) HSPCs. The HSPC apoptotic rate was detected by Ghost Dye™ Red 780 staining. The S-phase fraction significantly differed in various types of HSPCs. The values ranged from ≈ 1 % to ≈ 90 % of cells in S-phase, and were a characterizing feature of particular types of HSPCs. The CD71 (transferrin receptor) expression level closely correlated with the proliferation rate in all types of HSPCs. The highest S-phase fraction was found in the Lin c Kit+Sca-1- progenitors committed to the megakaryocyte erythroid development, followed by the progenitors committed to the granulocyte macrophage development. The HSPCs lacking the Sca-1 antigen generated approximately 22 times more cells than the Sca 1 expressing ones. Research was supported by the Grant Agency of the Czech Republic (GACR 17-01897S).

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