雪旺细胞无血清条件培养液中的神经营养蛋白活性

目的　进一步探测雪旺细胞无血清条件培养液中神经营养蛋白活性 .方法　收集成年大鼠坐骨神经雪旺细胞无血清条件培养液 (Schwanncellserum -freecondionedmedium ,SC -SFCM ) ,通过PM10超滤获取 >10Kd浓缩液 ,再经Disc -PAGE分离和Biotrap电洗脱 ,从SC -SFCM中分离出A、B、C、D四组蛋白带 ,选择其最佳活性浓度 ,四组蛋白带按照 7种不同组合加入脊髓前角神经元 (SAHN)培养液中 ,通过MTT法进行神经营养活性检测 .结果　含D蛋白带的各组 ,其OD值均显著高于对照组 p <0 .0 1) ,而不含D蛋白带的其它各组 ,其OD值与对照组比较无显著性差异 (p >0 .0 5 ) .结论　来自于SC -SFCM中的D蛋白带具有明显促进SAHN体外存活的神经营养活性

Activity of Neurotrophic Proteins Derived from Schwann Cell Serum-free Condioned Medium

Objective Further to explore activity of neurotrophic proteins derived from Schwann cell serum-free condioned medium(SC-SFCM). Methods SC-SFCM was collected and dialyzed in a ultrafiltration unit with Amicon PM10 membrane,recovering >10Kd concentrated fraction, which was then isolated and eluted by Disc-PAG and Biotrap apparatus, recovering A,B,C and D protein bands. In accordance with the best activity concentration of each protein band they were added into the spinal anterior horn neuron's medium by means of 7 different compositions. Neurotraphic activities were observed by MTT assay. Results Optical densite value of each composition containing D protein band was greatest than control(p<0.01),and no significant difference was observecd between each composition without D protein band and control (p>0.05). Conclutions This report suggests that D protein band derived from SC-SFCM has significant neurotrophic activity in vitro facilitating spinal anterior horn neuron survival.