Properties of primary murine stroma induced by macrophage colony-stimulating factor.
The ability of purified human macrophage colony-stimulating factor (M-CSF) to accelerate the formation of stromal cells from murine bone marrow cells was investigated. The liquid culture of the marrow cells with M-CSF resulted in the formation of monolayers of macrophages on day 7. When the M-CSF was removed on that day and the residual adherent cells were cultured in the absence of M-CSF for an additional 7 days, many colonies appeared with cells that were morphologically distinguishable from M-CSF-derived macrophages. The appearance of the colonies was dependent on the concentration of M-CSF used at the beginning of the culture. Each colony was isolated as a single clone and analyzed. All clones were negative for esterase staining. These cells did not express M-CSF receptor mRNA and did not show a mitogenic response to M-CSF. On the contrary, these cells could be stimulated to proliferate by fibroblast growth factor and platelet-derived growth factor. The polymerase chain reaction analysis of these cells demonstrated constitutive expression of mRNA for M-CSF, stem cell factor, and interleukin (IL)-1, but not IL-3. Some clones expressed mRNA for granulocyte/M-CSF and IL-6. We also examined the ability of the cells to maintain murine bone marrow high proliferative potential colony-forming cells (HPP-CFC) in a coculture system. Most of the clones showed a significant increase in total HPP-CFC numbers after 2 weeks of coculture, although the extent of stimulation differed among clones. These results suggested that the colonies established by M-CSF were composed of functional stromal cells that were phenotypically different from macrophages.
Yamada
M
M
Biochemical Research Laboratory, Morinaga Milk Industry Co. Ltd., Zama, Kanagawa, Japan.
Suzu
S
S
Akaiwa
E
E
Wakimoto
N
N
Hatake
K
K
Motoyoshi
K
K
Shimamura
S
S
D000818:Animals; D001854:Bone Marrow Cells; D002452:Cell Count; D002454:Cell Differentiation; D002478:Cells, Cultured; D002999:Clone Cells / Q000166:cytology / Q000378:metabolism; D018920:Coculture Techniques; D003114:Colony-Forming Units Assay; D004351:Drug Resistance; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005472:Fluorouracil / Q000494:pharmacology; D006133:Growth Substances / Q000235:genetics; D006651:Histocytochemistry; D006801:Humans; D016173:Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000494:pharmacology*; D008264:Macrophages / Q000166:cytology; D051379:Mice; D008815:Mice, Inbred Strains; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016186:Receptor, Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D017154:Stromal Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism
D006133:Growth Substances; D010982:Platelet-Derived Growth Factor; D012333:RNA, Messenger; D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2; D016173:Macrophage Colony-Stimulating Factor; D016186:Receptor, Macrophage Colony-Stimulating Factor; D005472:Fluorouracil