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.
New Drug Research Laboratories, Fujisawa Pharmaceutical Co., Ltd., Osaka, Japan.
Y
Y
Ueda
E
E
Walsh
H
H
Nakanishi
K
K
Yoshida
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