Expression of hematopoietic inhibitory factors in mouse fibroblasts, macrophages and endothelial cells.
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-1alpha, MIP-2, TGF-beta, TNF-alpha, IFN-gamma and Tbeta(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-alpha, INF-gamma and Tbeta(4), was seen in all three types of stromal cells, while TGF-beta mRNA was only seen in endothelial cells and macrophages, and MIP-1alpha mRNA in endothelial cells and fibroblasts. The inhibitors TGF-beta, MIP-1alpha, and Tbeta(4)have an inhibitory effect on the growth of CFU-GM, but TNF-alpha, INF-gamma and MIP-2 do not.
Cheng
La Mei
LM
Xiang Ya Medical School, Central South University, 410078 Changsha, China.
Yan
Yan
Y
Wang
Qi Ru
QR
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D017077:Culture Media, Conditioned / Q000494:pharmacology; D016207:Cytokines / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D004727:Endothelium / Q000166:cytology / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D015870:Gene Expression; D042381:Granulocyte Precursor Cells / Q000166:cytology / Q000187:drug effects; D006131:Growth Inhibitors / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D012333:RNA, Messenger / Q000378:metabolism; D017154:Stromal Cells / Q000166:cytology
D017077:Culture Media, Conditioned; D016207:Cytokines; D006131:Growth Inhibitors; D012333:RNA, Messenger