Bioassay detection of mouse nerve growth factor (mNGF) in the brain of adult mice

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Abstract
Two pools of seven brains each from adult Swiss‐Webster mice were homogenized, and supernatants were collected for bioassay. PC‐12 cells were placed in a bioassay plate at time zero, at a concentration of 104 cells per well, and primed for 48 hours in a medium containing 50 ng/ml of mNGF. The PC‐12 cell bioassay for neurite outgrowth was conducted after primed cells were exposed to an NGF‐free medium for 24 hours. Suitable controls for serum toxicity and cell viability were established. The sensitivity of the bioassay approximates 100 pg NGF/ml. The results showed 80–100% neurite outgrowth in wells exposed to brain pool supernatant (BPS) alone, and control level outgrowth (3–8%) in wells containing BPS and specific anti‐β‐NGF antibody. Therefore, the brains of Swiss‐Webster adult mice contain an NGF‐like substance which promotes neurite outgrowth in PC‐12 cells. The substance probably is NGF itself, since the effect is blocked by specific NGF antiserum.