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  • Title: Isolation and characterization of primitive hematopoietic progenitors of murine fetal liver.
    Author: Fujimoto K, Lyman SD, Hiryama F, Ogawa M.
    Journal: Exp Hematol; 1996 Feb; 24(2):285-90. PubMed ID: 8641354.
    Abstract:
    We have isolated hematopoietic progenitors from mouse fetal liver using a sequential protocol of density gradient centrifugation, panning, and cell sorting. Isolated AA4.1+Ly-6A/E+CD43++ cells with density ranging from 1.0631 to 1.0770 g/cm3 were 480- to 600-fold enriched for multipotential progenitors relative to unfractionated cells and showed 40 to 60% colony-forming efficiency. We then examined the effects of various cytokines on the colony formation from enriched fetal liver cells. Steel factor (SF), interleukin-3 (IL-3), IL-4, IL-6, IL-11, and granulocyte colony-stimulating factor (G-CSF) as single agents supported formation of significant numbers of colonies, but flt3/flk-2 ligand (FL) and IL-12 did not. When the cytokines were combined, FL, SF, IL-3, and IL-4 each synergized individually with IL-6, IL-11, IL-12, or G-CSF to support formation of various types of colonies. Next we analyzed the growth factor requirements for proliferation and differentiation of lymphohematopoietic progenitors by using the two-step methylcellulose culture assay we established recently. None of the early-acting factors were effective as a single agent, but combinations of SF or FL with IL-6, IL-11, or G-CSF were effective in supporting B cell potential of the primary colonies. Of these, the combination of FL plus IL-11 appeared to be the most effective.
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