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From the * Department of Molecular Genetics, Faculty of Medicine, Kyoto University, Kyoto 606-01, Japan; and the The establishment of culture conditions that selectively support hematopoietic stem cells is an
important goal of hematology. In this study, we investigated the possibility of using for this purpose a defined medium, mSFO2, which was developed for stromal cell-dependent bone
marrow cultures. We found that a combination of epidermal growth factor (EGF), the OP9
stromal cell line, which lacks macrophage colony-stimulating factor, recombinant stem cell factor, and the chemically defined medium mSFO2 provides a microenvironment where c-Kit+
Thy-1+/lo Mac-1+/lo B220 Institute of Allergy, Research Center Kyoto, Bayer Yakuhin, Ltd., Kyoto 619-02, Japan
TER119 common
+ IL-2R
+ gp130+ cells are selectively propagated from normal, unfractionated bone marrow cells. This cell population produced an in
vitro colony at a very high efficiency (50%), whereas it has only limited proliferative ability in
the irradiated recipient. Thus, the cells selected in this culture condition might represent colony-forming units in culture (CFU-c) with short-term reconstituting ability. Transferring this
cell population into medium containing differentiation signals resulted in the rapid production
of mature myelomonocytic and B cell lineages in vitro and in vivo. The fact that a similar culture condition was created by erb-B2-transduced OP9 in the absence of EGF indicated that
EGF exerts its effect by acting on OP9 rather than directly on CFU-c. These results suggested that the balance between self-renewal and differentiation of CFU-c can be regulated by extracellular signals.
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