title: Preferential proliferation of murine colony-forming units in culture in a chemically defined condition with a macrophage colony-stimulating factor-negative stromal cell clone.
abstract: 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- TER119- common beta + IL-2R gamma + 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 extra-cellular signals.
authors.affiliation: Department of Molecular Genetics, Faculty of Medicine, Kyoto University, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Takakura
authors.forename: H
authors.initials: H
authors.lastname: Kodama
authors.forename: S
authors.initials: S
authors.lastname: Nishikawa
authors.forename: S
authors.initials: S
authors.lastname: Nishikawa
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells; D002455:Cell Division / Q000187:drug effects; D002999:Clone Cells; D003114:Colony-Forming Units Assay; D017077:Culture Media, Conditioned; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects; D007700:Kinetics; D016173:Macrophage Colony-Stimulating Factor; D051379:Mice; D008810:Mice, Inbred C57BL; D011994:Recombinant Proteins / Q000494:pharmacology; D019089:Stem Cell Factor / Q000494:pharmacology; D017154:Stromal Cells / Q000166:cytology
chemical_list: D017077:Culture Media, Conditioned; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D016173:Macrophage Colony-Stimulating Factor