摘要
本文制备了巨噬细胞条件培养基(MφCM),并应用快速自动比色微量分析法检测MφCM对体外培养的生后7dSD大鼠小脑皮质神经元的作用。结果表明,分子量大于10kD的MφCM对神经元(细胞密度1×106/ml)的作用,与对照组比较有明显的神经营养活性(F<0.05);用盖玻片培养法表明,该组份有促进神经元突起生长的作用。MφCM经SephacrylS-100-HR凝胶层析和生物活性鉴定,获得了具有神经营养活性的第二峰洗脱液。此洗脱液经SDS-聚丙烯酰胺凝胶电泳分析,证明MφCM中神经营养活性成份的蛋白质其分子量为31kD-68kD之间。
n the present study,macrophage conditioned medium(M CM) was obtained, and the effect ofM CM on the survival and activity of cultured cerebellar cortical neurons of 7-day-old Spraque-Dawleyrat was measured by a rapid automated Colorimetric microassay. The data indicated that M CM frac-tion(>10kD) posseses significant neurotrophic activity to neurons at a density of 1×105 cells per millil-itre, as compared to the control (p<0. 01).Using coverslip culture method, above M CM also showsneurite promoting effects on the neurons. Additionally,the active fraction of M CM has been furtherpurified by Sephacryle S-100-HR gel chromatography and bioassay.Neurotrophic activity was found inthe M CM sephacryle-peak Ⅱ eluate。 This M CM sephacryle peak Ⅱ eluate was analysed by SDS-PAGE, The results identify that the molecular weight of neurotrophic active protein of M CM thatcould support the neuronal survival and enhance the neuronal activity is between 31kD and 68kD.
出处
《解剖科学进展》
CAS
1996年第4期363-369,共7页
Progress of Anatomical Sciences
基金
国家自然科学基金