Monday, May 03, 2004
Description Culture of Dissociated Sympathetic Neurons Procedure 1. Use day 9 chicken embryos. 2. Dissect sympathetic chains in Ham's F12 media. 3. Dissociate ganglia through incubation with 0.25% trypsin for 20 min at 37癈. 4. Wash tissue with 100% fetal calf serum for 5 minutes to end trypsinization 5. Wash chains 2x in Ham's F12 medium. 6. Dissociate tissue by gentle trituration with a flamed pipet 7. Plate onto pretreated (poly-dl-ornithine or poly-l-lysine) culture dishes containing 1.5 ml of either Neurobasal or serum-free F12+ medium 8. Add 10 ng/ml NGF Recipes F12+ Medium contains these supplements: - 100 礸/ml human transferrin - 1 ml Pen/Strept/100ml - 100 礛 putrescine - 20 nM progesterone - 30 nM selenium - 5 礸/ml bovine insulin Supplies Tips (责任编辑:泉水) |