Schwegler J S, Knorz M C, Akkoyun I, Liesenhoff H
University Eye Hospital, Klinikum Mannheim, D-68167 Mannheim, Germany.
Mol Vis. 1997 Oct 15;3:10.
In this study, we evaluated a possible effect of acidic and basic fibroblast growth factor (aFGF, bFGF) on the proliferation of human retinal pigment epithelial (RPE) cells in culture. As the RPE is the primary source for bFGF in the retina, such an effect would suggest autocrinic actions of FGFs.
Primary cultures of human and porcine RPE and an established human RPE cell line (D407) were subjected to aFGF and bFGF at different culture conditions. Cell proliferation was determined using the BrdU non-radioactive nucleotide analogue assay, and total protein was measured colorimetrically. The cells were subjected to aFGF and bFGF from 0.1 to 100 ng/ml for 1 to 14 days.
In the presence of 100 ng/ml bFGF, cell proliferation doubled from day 2 (143+/-12 units) to day 6 (227+/-17). This effect was neither seen without bFGF nor with aFGF at the same concentration. The stimulating effect of bFGF on cell proliferation was dose-dependent, the ED50 being around 1-10 ng/ml. The bFGF effect was markedly greater at high fetal calf serum concentration (10% vs. 1%). No bFGF effect was seen on cells of the established human RPE cell line D407 nor on primary cultures from porcine RPE.
bFGF, in contrast to its analogue aFGF, stimulates cell proliferation in cultured human RPE cells. It may act as an autocrinic agent (secretion by and action on the same cell) and thus be a specific regulator for cell proliferation in repair and replacement of the RPE cell monolayer.
在本研究中,我们评估了酸性和碱性成纤维细胞生长因子(aFGF、bFGF)对培养的人视网膜色素上皮(RPE)细胞增殖的可能影响。由于RPE是视网膜中bFGF的主要来源,这种影响将提示FGF的自分泌作用。
在不同培养条件下,将人及猪RPE的原代培养物和已建立的人RPE细胞系(D407)暴露于aFGF和bFGF。使用BrdU非放射性核苷酸类似物测定法确定细胞增殖,并通过比色法测量总蛋白。将细胞暴露于0.1至100 ng/ml的aFGF和bFGF中1至14天。
在存在100 ng/ml bFGF的情况下,细胞增殖从第2天(143±12单位)到第6天(227±17)翻倍。在没有bFGF的情况下或在相同浓度的aFGF存在下均未观察到这种效应。bFGF对细胞增殖的刺激作用是剂量依赖性的,ED50约为1-10 ng/ml。在高胎牛血清浓度(10%对1%)下,bFGF的作用明显更大。在已建立的人RPE细胞系D407的细胞或猪RPE的原代培养物中均未观察到bFGF的作用。
与类似物aFGF相比,bFGF可刺激培养的人RPE细胞增殖。它可能作为一种自分泌剂(由同一细胞分泌并作用于同一细胞),因此是RPE细胞单层修复和替换中细胞增殖的特异性调节剂。