Hoppenreijs V P, Pels E, Felten P C, Ruijter J M, Vrensen G F, Treffers W F
Department of Morphology, Netherlands Ophthalmic Research Institute, Amsterdam, The Netherlands.
Cornea. 1996 Jul;15(4):386-96. doi: 10.1097/00003226-199607000-00009.
Basic fibroblast growth factor (bFGF) is a major mitogen and chemoattractant for many cell types. The synergistic role of fetal bovine serum (FBS) and heparin on the modulation of tissue-cultured bovine corneal endothelial cells by bFGF was studied. Cell modulation was assessed by DNA synthesis ([3H]thymidine incorporation), mitochondrial activity (MTT assay), mitochondrial volume (transmission electron microscope study), and cell shape in mitotically competent cells and cells mitotically inhibited by 5-fluorouracil, mitomycin C, and irradiation. The dose-dependent stimulation of bFGF was found to be different for DNA synthesis and mitochondrial activity, the maximal effective dose being 1 and 10 ng/ml, respectively. Supplementation of bFGF with FBS enhanced both DNA synthesis and mitochondrial activity and caused a shift in the dose response to lower bFGF concentrations for the DAN synthesis, but not for the mitochondrial activity. Supplementation of bFGF with heparin resulted in an additional response for the mitochondrial activity, but not for the DNA synthesis. In addition, bFGF increased mitochondrial volume and induced elongation of the cells. These processes were further enhanced by the addition of heparin to bFGF. These results suggest that serum and heparin have a differential effect on mitogenic and non-mitogenic cells processes modulated by bFGF.
碱性成纤维细胞生长因子(bFGF)是多种细胞类型的主要促有丝分裂剂和趋化因子。研究了胎牛血清(FBS)和肝素对bFGF调节组织培养牛角膜内皮细胞的协同作用。通过DNA合成([3H]胸腺嘧啶核苷掺入)、线粒体活性(MTT法)、线粒体体积(透射电子显微镜研究)以及处于有丝分裂能力的细胞和被5-氟尿嘧啶、丝裂霉素C和辐射有丝分裂抑制的细胞的细胞形态来评估细胞调节。发现bFGF对DNA合成和线粒体活性的剂量依赖性刺激不同,最大有效剂量分别为1和10 ng/ml。bFGF与FBS联合使用可增强DNA合成和线粒体活性,并使DNA合成的剂量反应向较低的bFGF浓度偏移,但对线粒体活性无此影响。bFGF与肝素联合使用可使线粒体活性产生额外反应,但对DNA合成无此影响。此外,bFGF增加线粒体体积并诱导细胞伸长。向bFGF中添加肝素可进一步增强这些过程。这些结果表明,血清和肝素对bFGF调节的有丝分裂和非有丝分裂细胞过程具有不同的作用。