Ziats N P, Goldsmith G H, Medvedeff E D, Robertson A L
Thromb Res. 1986 Jan 15;41(2):239-50. doi: 10.1016/0049-3848(86)90232-x.
A new method for isolation and culture of endothelial cells from bovine coronary artery (BCoAEC) is presented. This method involves in situ perfusion and digestion of main coronary arteries with a collagenase solution. The isolated cells were cultured and maintained through many cell passages in Dulbecco's Modified Eagle's Medium supplemented with fetal bovine serum derived from either whole blood or plasma. Confirmation of these cells' endothelial origin was obtained by demonstration of typical morphologic and growth characteristics of endothelium, immunofluorescent staining with antibodies to von Willebrand factor (Factor VIII: vWF), and measurement of plasminogen activator (PA). In addition, production of PA was inhibited by enzymatically active thrombin as has been previously described with bovine aortic endothelial cells in culture.
本文介绍了一种从牛冠状动脉分离和培养内皮细胞（BCoAEC）的新方法。该方法包括用胶原酶溶液对主要冠状动脉进行原位灌注和消化。分离出的细胞在补充有全血或血浆来源的胎牛血清的杜氏改良 Eagle 培养基中培养，并通过多次传代维持。通过证明内皮细胞的典型形态和生长特征、用抗血管性血友病因子（因子 VIII：vWF）抗体进行免疫荧光染色以及测量纤溶酶原激活物（PA），证实了这些细胞的内皮来源。此外，如先前在培养的牛主动脉内皮细胞中所描述的那样，酶活性凝血酶抑制了 PA 的产生。
