CELL KINETICS IN DIFFUSION CHAMBERS: SURVIVAL, RESUMPTION OF PROLIFERATION, AND MATURATION RATE OF MURINE HAEMOPOIETIC CELLS

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Abstract
Bone marrow cells and blood leucocytes from mice have been cultured in vivo in diffusion chambers. Granulocytes and macrophages were formed in the chambers, whereas lymphocytes and mature end cells were gradually lost.The proliferation, differentiation and death of cultured cells were quantified by total and differential cell counting, by scintillation counting and by radio‐autographic evaluation of 3H‐thymidine incorporation. Hydroxyurea and vinblastine were used as cytotoxic agents.Ordinarily 40–60% of the cells inoculated could not be recovered after a few hours in culture. Proliferative cells resumed DNA synthesis shortly after implantation, and the rate of synthesis did not decline on further culturing. Steady‐state progenitor cells had a pre‐replicative lag period of about 18 hr, whereas regenerating progenitors started to synthesize DNA after about 14 hr. After 72 hr of culturing newly formed segmented granulocytes were detected. Maturation from the myelocyte to the segmented granulocyte stage lasted about 18 hr. Eosinophilic granulocytes had a long life span in the chambers, whilst the segmented neutrophils were rapidly eliminated after a life span estimated to be about 2 days.