Glassberg M K, Bern M M, Coughlin S R, Haudenschild C C, Hoyer L W, Antoniades H N, Zetter B R
In Vitro. 1982 Oct;18(10):859-66. doi: 10.1007/BF02796327.
Cells derived from the endothelium of human iliac arteries were cultured in vivo. The cells were isolated, grown, and subcultured in HEPES buffered Medium 199 supplemented with 20% heat inactivated human whole blood serum, human alpha-thrombin, and commercial endothelial cell growth supplement derived from bovine brain. The cells were viable in culture for 8 to 10 passages at a split ratio of 1:3. After the 10th passage, the cells began to enlarge and their growth rate was reduced. No cultures were viable after the 12th passage. The cells were determined to be of endothelial origin by their morphology at confluence; their ultrastructural characteristics, including the presence of Weibel-Palade bodies; the production and release of factor VIII-related antigen; and by their maintenance of a surface that prevented platelet attachment. The cultured arterial endothelial cells released prostacyclin in response to challenge with thrombin and protamine sulfate but not in response to bradykinin or the platelet-derived growth factor. Although the cultures described in this report were derived from patients with varying degrees of atherosclerotic disease, there were no significant differences in morphological or physiological parameters among these cultures or in comparison with commonly studied cells derived from human umbilical veins.
从人髂动脉内皮分离出的细胞在体内进行培养。这些细胞在补充有20%热灭活人全血血清、人α - 凝血酶和源自牛脑的商业内皮细胞生长补充剂的HEPES缓冲的199培养基中进行分离、培养和传代培养。细胞以1:3的传代比例在培养中可存活8至10代。第10代后,细胞开始增大且生长速率降低。第12代后无培养物存活。通过汇合时的形态、其超微结构特征(包括存在Weibel - Palade小体)、VIII因子相关抗原的产生和释放以及通过维持防止血小板附着的表面来确定细胞来源于内皮。培养的动脉内皮细胞在受到凝血酶和硫酸鱼精蛋白刺激时释放前列环素,但对缓激肽或血小板衍生生长因子刺激无反应。尽管本报告中描述的培养物来自患有不同程度动脉粥样硬化疾病的患者,但这些培养物之间以及与通常研究的源自人脐静脉的细胞相比,在形态或生理参数上没有显著差异。