Characterization of monocyte-macrophage-lineage cells induced from CD34+ bone marrow cells in vitro.
We characterized the expression of cell surface antigens and cytokine-secreting ability of monocyte-macrophage-lineage cells induced in vitro from CD34+ bone marrow cells. After cultivation for 3 weeks, we observed 2 distinct cell fractions: a floating small, round cell fraction and an adherent large, protruding cell fraction. Both cell fractions expressed myelocyte-monocyte-lineage antigens, but mature-macrophage markers such as CD206 were expressed only by the adherent cells. An assessment of cells cultured for 5 weeks revealed spontaneous secretion of interleukin 8 (IL-8) and IL-6, and lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNF-alpha) secretion in both fractions, but only the adherent cell fraction secreted IL-10 after LPS stimulation. In contrast, both fractions of cells cultured for 3 weeks spontaneously secreted low levels of IL-8, but none of the other cytokines. Upon LPS stimulation, the cells secreted IL-6 and TNF-alpha, but not IL-10. We also assessed the effect of granulocyte colony-stimulating factor (G-CSF) pretreatment on TNF-alpha secretion by each cell fraction and found that G-CSF reduced TNF-alpha secretion only in the adherent fraction of cells cultured for 3 weeks. Monocyte-macrophage-lineage cells induced in vitro should provide an ideal model for functional analysis of monocyte-macrophage cells.
Suzuki
Kyoko
K
Department of Developmental Biology, National Research Institute for Child Health and Development, Tokyo, Japan.
Kiyokawa
Nobutaka
N
Taguchi
Tomoko
T
Takenouchi
Hisami
H
Saito
Masahiro
M
Shimizu
Toshiaki
T
Okita
Hajime
H
Fujimoto
Junichiro
J
D018952:Antigens, CD34 / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology; D019070:Cell Lineage / Q000276:immunology; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism / Q000494:pharmacology; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D016130:Immunophenotyping; D008264:Macrophages / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D009000:Monocytes / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D010587:Phagocytosis / Q000276:immunology
D018952:Antigens, CD34; D016207:Cytokines; D016179:Granulocyte Colony-Stimulating Factor