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


CONCLUSIONS
MFB-F11 cells can be used to rapidly and specifically measure TGF-beta with high sensitivity.
Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Stanford, CA 94305, USA. itesseur@stanford.edu
Ina
I
Tesseur
Kun
K
Zou
Elisabeth
E
Berber
Hui
H
Zhang
Tony
T
Wyss-Coray
D000469:Alkaline Phosphatase / Q000235:genetics; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D001923:Brain Chemistry; D016466:CHO Cells; D002460:Cell Line; D018920:Coculture Techniques / Q000379:methods; D006224:Cricetinae; D003470:Culture Media / Q000737:chemistry; D005260:Female; D005347:Fibroblasts / Q000737:chemistry / Q000166:cytology; D017930:Genes, Reporter / Q000235:genetics; D051379:Mice; D008817:Mice, Mutant Strains; D012680:Sensitivity and Specificity; D015398:Signal Transduction / Q000502:physiology; D016212:Transforming Growth Factor beta / Q000032:analysis* / Q000097:blood / Q000235:genetics
D003470:Culture Media; D016212:Transforming Growth Factor beta; D000469:Alkaline Phosphatase