血管内皮细胞生长因子的制备和生物活性测定

PURIFICATION OF VASCULAR ENDOTHELIAL CELL GROWTH FACTOR AND DETERINATION OF ITS BIOLOGICAL ACTIVITY

报道从牛脑制备内皮细胞生长因子（ＥＣＧＦ）的方法。鲜牛脑经匀浆、硫酸铵盐析、透析后，上ＡｍｂｅｒｌｉｔｅＩＲＣ－５０离子交换柱，并用氯化钠梯度洗脱。利用ＥＣＧＦ对人脐静脉内皮细胞（ＨＵＶＥＣ）的增殖刺激效应测定其活性。当所得ＥＣＧＦ制品在含１０％新生牛血清或胎牛血清的培养液中含量为２５ｍｇ／Ｌ时，其最大刺激活性比对照组增加５５０％。

Purified endothelial cell growth factor(ECGF)was prepared from emulsified fresh bovine brain by fractionation (ammonium sultate precipitation method),chromatography on Amberlite IRC-50 and elution with double gradients containing 0～1 mol/L and 1～2mol/L NaCl. The eluates containing ECGF(10 μl each)were added to HUVEC culture to test the stimulating effect on cell proliferation by using colorimetric MTT assay.It was found that ECGF stimulated HUVEC proliferation in the presence of 10% NBS or FCS,HUVEC proliteration was a function of the ECGF concentration in culture medium, the estimated AC50 was 10 mg/L and the maximal stimulating effect of ECGF was found at the concentration of 25mg/L,where the proliferation was increased to 550%(5.5 times higher than the control).