Halle W, Sávoly B, Jakob W, Heder G, Baeger I
Acta Histochem Suppl. 1983;27:253-8.
A functionally and structurally well-defined calf aortic endothelial cell line has been used to characterize tissue preparations with mitogenic activity by means of cell proliferation kinetics. The following preparations were tested: An extract from corpora lutea (pig), produced by stepwise ammonium sulfate precipitation (CL-S3), a fraction obtained from CL-S3 extract by ion exchange chromatography (CM-S4), and a chorioallantoic membrane preparation (CAM preparation). Under optimal cultivation conditions 100 micrograms CL-S3, 5 micrograms CM-S4, or 100 micrograms CAM preparation per ml medium increased the cell number and growth rate of the endothelial cells in approximately like manner after an incubation period of 48 hours. Compared with CL-S3 these results demonstrate a 20 fold enrichment of the mitogenic activity in the CM-S4 fraction. Under suboptimal cultivation conditions (i.e. a low cell number at the seeding and a reduced nutritive quality) 100 micrograms CL-S3 per ml medium stimulated the cell proliferation up to 12 times at the end of a 6-days-cultivation-period. These results are discussed in connection with the use of these preparations in the angiogenesis research and with respect to the application of selected preparations in cell culture media in order to reduce the serum concentrations.
一种功能和结构明确的小牛主动脉内皮细胞系已被用于通过细胞增殖动力学来表征具有促有丝分裂活性的组织制剂。测试了以下制剂:通过逐步硫酸铵沉淀法制备的黄体(猪)提取物(CL-S3),通过离子交换色谱从CL-S3提取物中获得的一个组分(CM-S4),以及一种绒毛尿囊膜制剂(CAM制剂)。在最佳培养条件下,每毫升培养基中加入100微克CL-S3、5微克CM-S4或100微克CAM制剂,孵育48小时后,内皮细胞的细胞数量和生长速率以大致相同的方式增加。与CL-S3相比,这些结果表明CM-S4组分中的促有丝分裂活性提高了20倍。在次优培养条件下(即接种时细胞数量少且营养质量降低),每毫升培养基中加入100微克CL-S3在6天培养期结束时可将细胞增殖刺激多达12倍。结合这些制剂在血管生成研究中的应用以及为了降低血清浓度而在细胞培养基中应用选定制剂的情况对这些结果进行了讨论。