A novel primary culture method for rat choroidal epithelial cells.

Neurosciences (Riyadh, Saudi Arabia) Pub Date : 2013-01-01
Sheng-Li Huang, Xi-Jing He, Zong-Fang Li, Lu Yao, Wei Shi
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Abstract

Objective: To establish a method for the culture of primary choroidal epithelial cells.

Methods: This descriptive experimental study was carried out in Xi`an Jiaotong University, Xi`an, China from September 2009 to August 2012. Choroidal epithelial cells were isolated from the choroid plexus tissues of the lateral ventricles from neonatal rats (n=36). The tissues were dissociated into small cell aggregates by a mechanical method, and cultured on plastic culture dishes containing Dulbecco`s modified Eagle`s medium with 10% fetal bovine serum and 10 ng/ml epidermal growth factor at 37 degrees C in an incubator with 5% humidified carbon dioxide. The cultured cells were examined by phase contrast microscope, electron microscopy, and immunocytochemistry.

Results: The cells showed typical morphologic characteristics of epithelial phenotypes with a cobblestone appearance in monolayer 7-9 days post-seeding. The electron microscopy spotted typical choroidal epithelial cells with microvilli on the cytomembrane, organelles in the cytoplasm, and tight junctions welding 2 adjacent cells. They were positive against anti-transthyretin immunostaining.

Conclusion: This culture technique, which does not require complex equipment and operation skills, might be a simple and efficient method for obtaining choroidal epithelial cells in sufficient number and purity from mixed primary cultures of rat tissue.

一种新的大鼠脉络膜上皮细胞原代培养方法。
目的:建立培养原代脉络膜上皮细胞的方法。方法:本描述性实验研究于2009年9月至2012年8月在西安交通大学进行。从新生大鼠侧脑室脉络膜丛组织中分离脉络膜上皮细胞(n=36)。通过机械方法将组织分离成小细胞聚集体,并在含有含有10%胎牛血清和10 ng/ml表皮生长因子的Dulbecco 's modified Eagle 's培养基的塑料培养皿中,在37℃的培养箱中与5%的湿化二氧化碳培养。用相差显微镜、电镜和免疫细胞化学检查培养细胞。结果:培养后7 ~ 9 d,细胞表现出典型的上皮表型,单层呈鹅卵石状。电镜下可见典型的脉络膜上皮细胞,细胞膜上有微绒毛,细胞质中有细胞器,相邻细胞紧密连接。抗甲状腺素免疫染色阳性。结论:该培养技术不需要复杂的设备和操作技巧,可以从大鼠组织混合原代培养中获得足够数量和纯度的脉络膜上皮细胞,是一种简单有效的方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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