Lang I, Hoffmann C, Olip H, Pabst M A, Hahn T, Dohr G, Desoye G
Institute of Histology and Embryology and Clinic of Obstetrics and Gynaecology, Karl-Franzens University of Graz/Austria.
Cell Prolif. 2001 Jun;34(3):143-55. doi: 10.1046/j.1365-2184.2001.00205.x.
A variety of growth factors promote the complex multistep process of angiogenesis. The mitogenic activity of vascular endothelial growth factors (VEGFs) and placental growth factors (PlGFs), known as cytokines acting predominantly on endothelial cells, was tested on human umbilical vein endothelial cells (HUVEC) and microvascular endothelial cells (MIEC) and compared with the potency of the universally acting basic fibroblast growth factor (FGF-2). The cells were seeded at different cell numbers and incubated with various doses of growth factors for a period of 24-72 h in culture medium +/- serum. Proliferation was determined by measuring the optical density after staining the cells with the tetrazolium salt WST-1. VEGF121 and VEGF165 increased the number of HUVEC and MIEC at low and high seeding densities various doses and incubation times. The efficiency of FGF-2 was less pronounced at high seeding densities of the cells under serum-free conditions. PlGF-1 and PlGF-2 stimulated mitogenesis on HUVEC only at low cell numbers and after a short incubation time by 125 +/- 3% and 102 +/- 5% (P < 0.001), respectively. Longer incubation times with the lower seeding density in the absence of FCS did not induce a significant stimulatory effect of the PlGFs. MIEC responded stronger to all growth factors. In particular under serum free conditions, PlGF-1 and PlGF-2 effectively stimulated cell proliferation by 247 +/- 54% (P < 0.01) and 288 +/- 40% (P < 0.05) at low cell numbers, and by 81 +/- 13% (P < 0.05) and 49 +/- 13% (P < 0.01), respectively, at high cell numbers. The addition of fetal calf serum caused a reduced proliferative response of all growth factors on both cell types related to the controls. In conclusion, MIEC and HUVEC differ in their proliferative response to VEGFs, PlGFs and FGF-2.
多种生长因子可促进血管生成这一复杂的多步骤过程。血管内皮生长因子（VEGF）和胎盘生长因子（PlGF）具有促有丝分裂活性，它们被认为是主要作用于内皮细胞的细胞因子。本研究在人脐静脉内皮细胞（HUVEC）和微血管内皮细胞（MIEC）上测试了这些因子的活性，并与普遍起作用的碱性成纤维细胞生长因子（FGF-2）的效力进行了比较。将细胞以不同数量接种，在含或不含血清的培养基中，与不同剂量的生长因子一起孵育24至72小时。通过用四唑盐WST-1对细胞染色后测量光密度来确定细胞增殖情况。VEGF121和VEGF165在不同接种密度、不同剂量和孵育时间下均增加了HUVEC和MIEC的数量。在无血清条件下，当细胞接种密度较高时，FGF-2的效力不太明显。PlGF-1和PlGF-2仅在低细胞数量且短时间孵育后才刺激HUVEC的有丝分裂，分别增加了125±3%和102±5%（P<0.001）。在无胎牛血清（FCS）且接种密度较低的情况下延长孵育时间，PlGFs并未产生显著的刺激作用。MIEC对所有生长因子的反应更强。特别是在无血清条件下，PlGF-1和PlGF-2在低细胞数量时分别有效刺激细胞增殖247±54%（P<0.01）和288±40%（P<0.05），在高细胞数量时分别刺激81±13%（P<0.05）和49±13%（P<0.01）。添加胎牛血清后，与对照组相比，所有生长因子对两种细胞类型的增殖反应均降低。总之，MIEC和HUVEC对VEGFs、PlGFs和FGF-2的增殖反应存在差异。
