A colorimetric assay method for the evaluation of neurotrophic activity in vitro.
A colorimetric assay was established to detect neurotrophic activity by measuring the lysosomal enzyme, acid phosphatase (AP) activity of cultured neuronal cells. Neurons from the cerebral cortex of 14- or 15-day mouse embryo were cultured in serum-free medium for 3 days in 96-well culture plates. A linear relationship was obtained between the AP activity and the number of viable neurons counted under a microscope. The AP assay was used to evaluate the neurotrophic activity of basic fibroblast growth factor. This assay is shown to be simple, sensitive and convenient to detect neurotrophic activity.
Ueda
Y
Y
New Drug Research Laboratories, Fujisawa Pharmaceutical Co., Ltd., Osaka, Japan.
Walsh
E
E
Nakanishi
H
H
Yoshida
K
K
D000135:Acid Phosphatase / Q000032:analysis; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003124:Colorimetry; D016895:Culture Media, Serum-Free; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D051379:Mice; D008815:Mice, Inbred Strains; D009474:Neurons / Q000187:drug effects* / Q000201:enzymology; D013778:Tetrazolium Salts; D013844:Thiazoles
D016895:Culture Media, Serum-Free; D013778:Tetrazolium Salts; D013844:Thiazoles; D016222:Fibroblast Growth Factor 2; D000135:Acid Phosphatase; C022616:thiazolyl blue