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  • Title: Rapid differentiation of a rare subset of adult human lin(-)CD34(-)CD38(-) cells stimulated by multiple growth factors in vitro.
    Author: Fujisaki T, Berger MG, Rose-John S, Eaves CJ.
    Journal: Blood; 1999 Sep 15; 94(6):1926-32. PubMed ID: 10477721.
    Abstract:
    Recently, several reports of lineage-negative (lin(-)) CD34(-) cells with in vivo hematopoietic activity have focused interest on the properties and growth factor response characteristics of these cells. We have now identified a combination of 5 growth factors that are necessary and sufficient to stimulate a marked mitogenic and differentiation response by a subset of human lin(-)CD34(-)CD38(-) cells present in normal adult human marrow and granulocyte colony-stimulating factor (G-CSF)-mobilized blood. Less than 0.1% of the cells in highly purified (including doubly sorted) lin(-)CD34(-)CD38(-) cells from these 2 sources formed colonies directly in semisolid medium or generated such cells after 6 weeks in long-term culture. Nevertheless, approximately 1% of the same lin(-)CD34(-)CD38(-) cells were able to proliferate rapidly in serum-free liquid suspension cultures containing human flt-3 ligand, Steel factor, thrombopoietin, interleukin-3 (IL-3), and hyper-IL-6 to produce a net 28- +/- 8-fold increase in total cells within 10 days. Of the cells present in these 10-day cultures, 5% +/- 2% were CD34(+) and 2.5% +/- 0.9% were erythroid, granulopoietic, megakaryocytopoietic, or multilineage colony-forming cells (CFC) (13 +/- 7 CFC per lin(-)CD34(-)CD38(-) pre-CFC). In contrast to lin(-)CD34(+)CD38(-) cells, this response of lin(-)CD34(-)CD38(-) cells required exposure to all of the 5 growth factors used. Up to 1.7 x 10(5) lin(-)CD34(-) adult marrow cells failed to engraft sublethally irradiated NOD/SCID-beta(2)M(-/-) mice. These studies demonstrate unique properties of a rare subset of lin(-)CD34(-)CD38(-) cells present in both adult human marrow and mobilized blood samples that allow their rapid proliferation and differentiation in vitro within an overall period of 3 to 4 weeks. The rapidity of this response challenges current concepts about the normal duration and coordinated control of these processes in adults.
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