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BMC Cell Biol. 2006 Mar 20;7:15.

Highly sensitive and specific bioassay for measuring bioactive TGF-beta.

Author information

  • 1Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Stanford, CA 94305, USA. itesseur@stanford.edu

Abstract

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.

PMID:
16549026
[PubMed - indexed for MEDLINE]
PMCID:
PMC1479809
Free PMC Article
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