Dömötör E, Sipos I, Kittel A, Abbott N J, Adam-Vizi V
Department of Medical Biochemistry, Semmelweis University of Medicine, Budapest, Hungary.
Neurochem Int. 1998 Dec;33(6):473-8. doi: 10.1016/s0197-0186(98)00057-6.
An improved method for culturing primary rat brain capillary endothelial cells on glass has been developed, using a corneal extracellular matrix coat. Since the collagen-coated plastic attachment surface conventionally used for primary cultures of brain microvascular endothelium gives a high level of background fluorescence in microfluorimetric studies, an alternative attachment surface was tested involving no plastic element. Five substrata combinations were examined and a new combination of glass and corneal endothelial extracellular matrix coat was found to provide excellent cell adhesion, culture growth and purity. Other established substrata combinations tested for comparison, either involved plastic, or used glass with collagen or carbodiimide and collagen coating but the last two gave poor endothelial cell adhesion and growth. Our method using this new attachment surface combination results in stable and pure endothelial cultures, as verified by immunocytochemistry, which are suitable for fluorimetric investigations.
已经开发出一种改进的方法，利用角膜细胞外基质包被在玻璃上培养原代大鼠脑毛细血管内皮细胞。由于传统上用于脑微血管内皮细胞原代培养的胶原包被塑料附着表面在微量荧光研究中会产生高水平的背景荧光，因此测试了一种不含塑料成分的替代附着表面。研究了五种基质组合，发现玻璃与角膜内皮细胞外基质包被的新组合能提供优异的细胞黏附、培养生长和纯度。为作比较而测试的其他既定基质组合，要么涉及塑料，要么使用涂有胶原或碳二亚胺与胶原的玻璃，但后两者的内皮细胞黏附和生长情况较差。我们使用这种新附着表面组合的方法可产生稳定且纯净的内皮细胞培养物，免疫细胞化学验证其适用于荧光研究。
