Characterization of Conditioned Media Collected from Aged versus Young Human Eye Cups
Characterization of Conditioned Media Collected from Aged versus Young Human Eye Cups
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DOI：
10.1167/iovs.10-6440
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发表时间：
2011-07-01
影响因子：
4.4
通讯作者：
Zarbin, Marco A.
中科院分区：
文献类型：
--
作者：
Kolomeyer, Anton M.;Sugino, Ilene K.;Zarbin, Marco A.
目的。目的研究健康、老年、老年性黄斑变性(AMD)成人和胎儿供眼的视网膜色素上皮(RPE)分泌特征，并评价其对视网膜存活的影响。方法：采集成人和胎儿供眼的条件培养液(CM)，用多重酶联免疫吸附试验(ELISA)分析其营养因子的组成。计算营养因子受体占有率以评估营养因子浓度的差异。实时定量聚合酶链式反应(Real-Time-PCR)检测RPE营养因子基因表达。结果与成年供体CM相比，AMD供体CM中脑源性神经营养因子(BDNF)含量显著升高，而胎儿供体CM中肝细胞生长因子(HGF)和色素上皮衍生因子(PEDF)含量显著升高。营养因子mRNA的表达与蛋白质分泌之间没有一致的相关性。RPE-Bruch膜-脉络膜-巩膜复合体中的非RPE成分是血管内皮生长因子-A(VEGF-A)的主要成分。胎儿供体的CM在提高变性猪视网膜存活率方面明显优于成人或AMD供体。结论成人和胎儿RPE细胞具有明显不同的营养因子产生能力，这种差异与猪视网膜保存的变化有关。结合营养因子受体占有率的计算，这些数据可能暗示HGF和PEDF是促进视网膜结构和功能保存的关键因素。(投资眼科VS科学。2011年；52：5963-5972)doi：10.1167/iovs.10-6440
PURPOSE. To characterize secretion of in situ retinal pigment epithelium (RPE) from healthy, aged adult, age-related macular degeneration (AMD) adult, and fetal donor eyes and to assess the impact on retinal survival in vitro.METHODS. Conditioned medium (CM) was collected from adult and fetal donor eyes and analyzed for trophic factor composition by multiplex ELISA. Trophic factor receptor occupancy was calculated to evaluate differences in trophic factor concentrations. RPE trophic factor mRNA expression was quantified by real-time PCR. Retina-preserving activity of the collected CM was evaluated using degenerating porcine retina in vitro.RESULTS. Compared with CM from adult donors, AMD donor CM contained a significantly higher concentration of brain-derived neurotrophic factor (BDNF), whereas fetal donor CM contained significantly higher concentrations of hepatocyte growth factor (HGF) and pigment epithelium-derived factor (PEDF). No consistent correlation was found between trophic factor mRNA expression and protein secretion. Non-RPE components of the RPE-Bruch's membrane-choroid-sclera complex were major contributors of vascular endothelial growth factor-A (VEGF-A). CM of fetal donors was significantly better than CM of adult or AMD donors at improving the survival of degenerating porcine retina.CONCLUSIONS. RPE cells of adult and fetal eyes have significantly different trophic factor production capabilities, which correlated with changes in preservation of porcine retina. Combined with trophic factor receptor occupancy calculations, these data may implicate HGF and PEDF as key factors promoting the preservation of retinal structure and function. (Invest Ophthalmol Vis Sci. 2011;52:5963-5972) DOI: 10.1167/iovs.10-6440