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首页> 外文期刊>Cell and Tissue Research >Experimental lens capsular bag model for posterior capsule opacification
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Experimental lens capsular bag model for posterior capsule opacification

机译：后囊混浊的实验性晶状体囊袋模型

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An in vitro culture model enabling posterior capsule opacification (PCO) to be investigated was developed and established by using low-melting-point (LMP)-agarose gel to support the capsular bag. After removal of the cornea from rodent and porcine eyeballs, the lens zonules were dissected. Whole lens explants were embedded into 2 % (37°C) LMP-agarose gel solution. As performed routinely in cataract surgery, capsulotomy and lens fiber removal were carried out in the solidified LMP-agarose gel as sham cataract surgery. The LMP-agarose-gel-supported capsular bag/lens epithelial cell (CB-LEC) complexes were maintained in Dulbecco's modified Eagle medium supplemented with 10 % fetal bovine serum in an anterior face-down position. The proliferation and migration of LECs into the posterior capsule were observed every 12 h by phase-contrast microscopy. Epithelial cells were observed at the central portion of the CB-LEC complexes after 56.57±16.56 h (n=7) and 106±14.03 h (n=6) of culture, for rodent and porcine lenses, respectively. The solidified gel allowed clear microscopic observations and whole-mount immunostaining evaluations of the whole area of the capsular bag. Histological examinations revealed the proliferation, migration, and transdifferentiation of LECs related to posterior capsule opacification. This new in vitro culture model provides experimental benefits by maintaining the natural contour of the capsule without implants inside or outside of the capsule. In addition, this model system allows pharmacological and histological evaluations of the cultured CB-LEC complexes without additional manipulations.
机译：通过使用低熔点（LMP）-琼脂糖凝胶支持囊袋，开发并建立了能够研究后囊混浊（PCO）的体外培养模型。从啮齿动物和猪眼球中去除角膜后，将晶状体小带切开。将整个晶状外植体包埋在2％（37°C）LMP-琼脂糖凝胶溶液中。如在白内障手术中常规进行的，作为假性白内障手术，在固化的LMP-琼脂糖凝胶中进行囊切开术和晶状体纤维去除。 LMP-琼脂糖凝胶支持的囊袋/晶状体上皮细胞（CB-LEC）复合物维持在Dulbecco改良的Eagle培养基中，该培养基添加了10％的胎牛血清，面朝下。每12小时通过相差显微镜观察LEC的增殖和迁移到后囊中。培养56.57±16.56h（n = 7）和106±14.03h（n = 6）后，分别在啮齿类和猪晶状体上观察到CB-LEC复合体中心部分的上皮细胞。固化后的凝胶可以清晰地观察到显微镜，并对囊袋的整个区域进行整体免疫染色评估。组织学检查显示与后囊混浊有关的LEC的增殖，迁移和转分化。这种新的体外培养模型通过保持胶囊的自然轮廓而无需在胶囊内部或外部植入植入物，从而提供了实验优势。此外，该模型系统无需额外操作即可对培养的CB-LEC复合物进行药理和组织学评估。

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