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Expression of hematopoietic inhibitory factors in mouse fibroblasts, macrophages and endothelial cells
Author(s) -
Cheng La Mei,
Yan Yan,
Wang Qi Ru
Publication year - 2003
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1016/s1065-6995(03)00144-6
Subject(s) - microbiology and biotechnology , haematopoiesis , cytokine , endothelial stem cell , cell culture , biology , bone marrow , fibroblast , stromal cell , colony forming unit , macrophage , chemistry , in vitro , immunology , biochemistry , stem cell , genetics , cancer research , bacteria
Pure bone marrow fibroblasts, macrophages and endothelial cells were cultured in Iscove‐modified Dulbecco's medium. RT—PCR was used to determine the expression of inhibitory cytokine mRNAs in these cell types. Serum‐free conditioned medium was collected from each cell type and ultrafiltration was performed with a centriprep 10. The retentate contained substances whose molecular weights were >10 kD, whilst the filtrate contained substances with molecular weights <10 kD. The effect of conditioned media and their components on colony forming unit‐granulocyte‐macrophage (CFU‐GM) were investigated. The results showed: (1) six cytokines, MIP‐1α, MIP‐2, TGF‐β, TNF‐α, IFN‐γ and Tβ 4 , inhibited the growth of CFU‐GM when murine WEHI‐3 conditioned medium was added to the culture system as a source of colony stimulation. (2) The original endothelial cell conditioned medium (E‐CM) did not affect the production of CFU‐GM, but the >10 kD component of E‐CM increased its production, and the <10 kD component decreased it. Both fibroblast conditioned medium (F‐CM) and the >10 kD component of F‐CM stimulated proliferation of CFU‐GM, but the <10 kD component suppressed it. All three components of macrophage conditioned medium (M‐CM) inhibited the growth of CFU‐GM. (3) Expression of four of the mRNAs, namely MIP‐2, TNF‐α, INF‐γ and Tβ 4 , was seen in all three types of stromal cells, while TGF‐β mRNA was only seen in endothelial cells and macrophages, and MIP‐1α mRNA in endothelial cells and fibroblasts. The inhibitors TGF‐β, MIP‐1α, and Tβ 4 have an inhibitory effect on the growth of CFU‐GM, but TNF‐α, INF‐γ and MIP‐2 do not.