Macarak E J, Howard B V, Kefalides N A
Lab Invest. 1977 Jan;36(1):62-7.
Calf aorta endothelial cells, obtained by collagenase treatment of vessels from freshly killed animals, were cultured in medium 199 supplemented with fetal bovine serum, amino acids, and vitamins. The material released from the vessel wall after collagenase treatment consists of clumps of cells which quickly attach to the dish and spread to form confluent islands of cells. These islands of cells coalesce to form a confluent monolayer in 6 to 8 days. Cultures labeled with [3H]thymidine show an increase in labeling commencing 3 days after initial culture. Cells within monolayers do not overgrow one another and retain their epithelioid appearance after subculture. Identification of endothelial cells was based on culture morphology and by the presence of factor VIII antigen as localized by indirect immunofluorescence microscopy. These data indicate that large numbers of endothelial cells (5 to 7 x 10(6) cells per aorta) can be obtained and maintained in culture.
通过用胶原酶处理刚处死动物的血管获得的小牛主动脉内皮细胞，在补充有胎牛血清、氨基酸和维生素的199培养基中培养。胶原酶处理后从血管壁释放的物质由细胞团组成，这些细胞团迅速附着于培养皿并铺展形成汇合的细胞岛。这些细胞岛在6至8天内融合形成汇合的单层。用[3H]胸腺嘧啶核苷标记的培养物在初始培养3天后显示标记增加。单层内的细胞不会相互过度生长，传代培养后仍保持其上皮样外观。内皮细胞的鉴定基于培养形态以及通过间接免疫荧光显微镜定位的因子VIII抗原的存在。这些数据表明，可以获得大量内皮细胞（每条主动脉5至7×10⁶个细胞）并在培养中维持。
