Sakamoto T, Sakamoto H, Hinton D R, Spee C, Ishibashi T, Ryan S J
Doheny Eye Institute, Los Angeles, CA 90033, USA.
Curr Eye Res. 1995 Aug;14(8):621-7. doi: 10.3109/02713689508998488.
Vascular endothelial cells play an important role in progression or healing of various retinal or choroidal diseases and they have a broad organ specificity. We have grown and studied human choroidal endothelial cells (CECs) cultured in a collagen gel, where they form tube-like structures. CECs were differentially isolated from choroidal tissues and cultured to near homogeneity. The cells were embedded in a type I collagen gel in 24-well culture dishes and incubated with M199 medium containing 10% fetal calf serum. Gels were observed using phase contrast microscopy and were evaluated histologically by light and electron microscopy. The CECs were strongly positive for factor VIII-related antigen and actively ingested diI-acetylated LDL, indicating their endothelial nature. Ultrastructural analysis of the tube-like structures revealed a central lumen surrounded by cells joined apically by junctions and showing prominent pinocytotic activity, fenestrations and basement membrane formation; these features are typical of CECs in vivo. This preparation should be a useful tool to study the development of physiology and pathology of choroidal endothelium.
血管内皮细胞在各种视网膜或脉络膜疾病的进展或愈合过程中发挥着重要作用，并且具有广泛的器官特异性。我们已经培养并研究了在胶原凝胶中培养的人脉络膜内皮细胞（CEC），它们在其中形成管状结构。CEC从脉络膜组织中差异分离并培养至接近均一性。将细胞嵌入24孔培养板中的I型胶原凝胶中，并用含有10%胎牛血清的M199培养基孵育。使用相差显微镜观察凝胶，并通过光镜和电镜进行组织学评估。CEC对VIII因子相关抗原呈强阳性，并能主动摄取DiI-乙酰化低密度脂蛋白，表明其内皮细胞性质。对管状结构的超微结构分析显示，中央管腔被通过顶端连接相连的细胞包围，并表现出明显的胞饮活性、窗孔和基底膜形成；这些特征是体内CEC的典型特征。这种制备方法应该是研究脉络膜内皮细胞生理和病理发育的有用工具。
