Schwartz S M
In Vitro. 1978 Dec;14(12):966-80. doi: 10.1007/BF02616210.
This paper reports techniques for isolation, selection and long-term passage of bovine aortic endothelium (BAE). A [3H]thymidine-selection technique was developed to limit overgrowth of cultures by contaminating smooth-muscle cells. The resulting cultures could be passaged for a replicative life span of 35 to 40 doublings and maintained a stable, normal karyotpye throughout this period. Despite the fact that these cultures reached a stable monolayer with density-inhibited growth state, postconfluent cells showed focal areas of a second growth pattern called "sprouting." This was seen only when cultures were maintained at high densities for periods of 1 to 2 weeks. Ultrastructural analysis, as well as immunofluorescence studies with markers for endothelial cells (factor VIII) and smooth-muscle cells (actin), indicates that this phenomenon is not due to overgrowth of a residual population of smooth-muscle cells, but may represent a second growth pattern of the endothelial cells themselves.
本文报道了牛主动脉内皮细胞（BAE）的分离、筛选及长期传代技术。开发了一种[3H]胸腺嘧啶核苷筛选技术，以限制培养物被平滑肌细胞污染而过度生长。由此得到的培养物可传代35至40次倍增的复制寿命，并在此期间保持稳定、正常的核型。尽管这些培养物达到了具有密度抑制生长状态的稳定单层，但汇合后细胞显示出一种称为“发芽”的第二种生长模式的局灶区域。仅当培养物在高密度下维持1至2周时才会出现这种情况。超微结构分析以及用内皮细胞（因子VIII）和平滑肌细胞（肌动蛋白）标记物进行的免疫荧光研究表明，这种现象不是由于平滑肌细胞残留群体的过度生长，而是可能代表内皮细胞自身的第二种生长模式。
