Highly sensitive and specific bioassay for measuring bioactive TGF-β
1 Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Stanford, CA 94305, USA
2 GRECC, VA Palo Alto Health Care System, Palo Alto, CA 94304, USA
3 Department of Alzheimer's Disease Research, National Center for Geriatrics and Gerontology, Obu, Aichi 474-8522, Japan
BMC Cell Biology 2006, 7:15 doi:10.1186/1471-2121-7-15Published: 20 March 2006
Transforming Growth Factor-β (TGF-β) regulates key biological processes during development and in adult tissues and has been implicated in many diseases. To study the biological functions of TGF-β, sensitive, specific, and convenient bioassays are necessary. Here we describe a new cell-based bioassay that fulfills these requirements.
Embryonic fibroblasts from Tgfb1-/- mice were stably transfected with a reporter plasmid consisting of TGF-β responsive Smad-binding elements coupled to a secreted alkaline phosphatase reporter gene (SBE-SEAP). Clone MFB-F11 showed more than 1000-fold induction after stimulation with 1 ng/ml TGF-β1, and detected as little as 1 pg/ml TGF-β1. MFB-F11 cells were highly induced by TGF-β1, TGF-β2 and TGF-β3, but did not show induction with related family members activin, nodal, BMP-2 and BMP-6 or with trophic factors bFGF and BDNF. MFB-F11 cells can detect and quantify TGF-β in biological samples without prior enrichment of TGF-βs, and can detect biologically activated TGF-β in a cell co-culture system.
MFB-F11 cells can be used to rapidly and specifically measure TGF-β with high sensitivity.