Highly sensitive and specific bioassay for measuring bioactive TGF-beta

BMC Cell Biol. 2006 Mar 20;7:15. doi: 10.1186/1471-2121-7-15.


Background: Transforming Growth Factor-beta (TGF-beta) regulates key biological processes during development and in adult tissues and has been implicated in many diseases. To study the biological functions of TGF-beta, sensitive, specific, and convenient bioassays are necessary. Here we describe a new cell-based bioassay that fulfills these requirements.

Results: Embryonic fibroblasts from Tgfb1-/- mice were stably transfected with a reporter plasmid consisting of TGF-beta 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-beta1, and detected as little as 1 pg/ml TGF-beta1. MFB-F11 cells were highly induced by TGF-beta1, TGF-beta2 and TGF-beta3, 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-beta in biological samples without prior enrichment of TGF-betas, and can detect biologically activated TGF-beta in a cell co-culture system.

Conclusion: MFB-F11 cells can be used to rapidly and specifically measure TGF-beta with high sensitivity.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / genetics
  • Animals
  • Biological Assay / methods*
  • Brain Chemistry
  • CHO Cells
  • Cell Line
  • Coculture Techniques / methods
  • Cricetinae
  • Culture Media / chemistry
  • Female
  • Fibroblasts / chemistry
  • Fibroblasts / cytology
  • Genes, Reporter / genetics
  • Mice
  • Mice, Mutant Strains
  • Sensitivity and Specificity
  • Signal Transduction / physiology
  • Transforming Growth Factor beta / analysis*
  • Transforming Growth Factor beta / blood
  • Transforming Growth Factor beta / genetics


  • Culture Media
  • Transforming Growth Factor beta
  • Alkaline Phosphatase