Human lens epithelial cell proliferation in a protein-free medium.
Human lens epithelial cell proliferation in a protein-free medium.
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DOI：
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发表时间：
1997-02
影响因子：
4.4
通讯作者：
M. Wormstone;C. ChristopherS.;Liu;Jean-Marie Rakic;Julia;-M.;Marcantonio;J. GijsF.;Vrensen;George Duncan
中科院分区：
文献类型：
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作者：
M. Wormstone;C. ChristopherS.;Liu;Jean-Marie Rakic;Julia;-M.;Marcantonio;J. GijsF.;Vrensen;George Duncan
目的：白内障囊外摘出术（ECCE）后，人眼内的体液蛋白含量相对较低，但囊袋内仍有细胞生长。这种弹性生长在很大比例（30%）的患者中引起后囊膜混浊（PCO）。本研究比较了人晶状体细胞在补充血清和无蛋白培养基中的增殖能力。方法对人供眼进行假白内障手术。分离荚膜袋，平钉在培养皿上，单独在Eagle最小必需培养基（EMEM）中或在添加10%胎牛血清的EMEM中孵育。用相衬显微镜观察。在终点，用荧光或电子显微镜对胶囊进行研究。用溴-2-脱氧尿苷标记和检测技术鉴定有丝分裂活性。必要时，手术时植入人工晶状体。结果发现，在没有添加蛋白的情况下，来自广泛年龄谱的供体的人晶状体细胞在晶状体囊上增殖和迁移。生长速度与年龄有关，60岁供体晶状体的后囊在8.0 +/- 0天（n = 3）和24.4 +/- 5.3天（n = 3）后完全融合。上皮细胞的生长引起包膜收缩、起皱和光散射增加。人工晶状体前表面的生长比后囊少。结论无蛋白模型复制了临床观察到的PCO的许多特征。天然胶原蛋白胶囊上弹性细胞的生长解释了PCO的高患病率，特别是在年轻患者中，并表明炎症和外部生长因子不是PCO的必要条件。此外，无蛋白荚膜袋系统可用于探索晶状体上皮细胞存活和生长的自分泌控制的基本问题。
PURPOSE The ocular humors are relatively low in protein, yet cell growth in the human capsular bag still occurs after extracapsular cataract extraction (ECCE) surgery. This resilient growth gives rise to posterior capsule opacification (PCO) in a significant proportion (30%) of patients. This study compared the ability of human lens cells to proliferate in serum-supplemented and protein-free medium. METHODS Sham cataract operations were performed on human donor eyes. The capsular bag was dissected free, pinned flat on a petri dish, and incubated in Eagle's minimal essential medium (EMEM) alone or in EMEM supplemented with 10% fetal calf serum. Observations were made by phase-contrast microscopy. At the endpoint, capsules were studied by fluorescence or electron microscopy. Mitotic activity was identified using Bromo-2-deoxyuridine labeling and detection techniques. When required, an intraocular lens was implanted when surgery was performed. RESULTS It was found that human lens cells from a wide age spectrum of donors proliferate and migrate on the lens capsule in the absence of added protein. The rate of growth was age-dependent, such that the posterior capsule was completely confluent after 8.0 +/- 0 days (n = 3) and 24.4 +/- 5.3 days (n = 3) for donor lenses aged 60 years, respectively. The outgrowth of epithelial cells gave rise to capsular contraction, wrinkling, and increased light scatter. Growth on the anterior surface of the intraocular lens was less prolific than on the posterior capsule. CONCLUSION The protein-free model replicates many features of clinically-observed PCO. The resilient cell growth on the natural collagen capsule explains the high prevalence of PCO, especially in younger patients, and suggests that inflammation and external growth factors are not necessary for PCO. Furthermore, the protein-free capsular bag system can be used to explore fundamental questions concerning the autocrine control of lens epithelial cell survival and growth.