Complex cytokine modulation of a continuous line of mink lung epithelial cells (Mv1Lu).
The continuous mink lung epithelial cell line Mv1Lu has proven to be a sensitive reporter line in the bioassay for purified TGF-beta, exhibiting a sigmoid-shaped concentration-response relationship with an EC50 of 12 pM (0.3 ng/mL). Maximal inhibition of Mv1Lu cells generates a 75-95% decrement in the number of adherent cells. However, this bioassay is not specific for TGF-beta as originally claimed. Mv1Lu cells are sensitive to other cytokines and substances found in complex biological fluids. In this study the effects of other biological response modifiers in this assay were tested and several were found to have important growth modulatory capacities that confound the quantitation of TGF-beta. EGF, TGF-alpha, fibronectin, and IGF-I all induce Mv1Lu cell proliferation. In contrast, neither PDGF (-AA, -AB, -BB) nor endotoxin (< or = 10 micrograms/mL) affect Mv1Lu cell number. TGF-beta and TNF-alpha at high concentrations (> or = 10 ng/mL) are the only cytokines examined that inhibit Mv1Lu proliferation. TGF-beta decreases final cell number both by preventing mitosis and by inhibition of adherence of cells to the uncoated dish. Several strategies are suggested to assure the specificity of this otherwise convenient bioassay for TGF-beta.
Kelley
J
J
Department of Medicine, University of Vermont College of Medicine, Burlington 05405.
Baldor
L
L
Absher
M
M
D000818:Animals; D001681:Biological Assay; D002452:Cell Count; D002460:Cell Line; D003124:Colorimetry; D016207:Cytokines / Q000502:physiology*; D004847:Epithelial Cells; D004848:Epithelium / Q000502:physiology; D008168:Lung / Q000166:cytology* / Q000502:physiology; D008907:Mink / Q000502:physiology*
D016207:Cytokines