论文部分内容阅读
随着人造血管内皮化研究的深入,需要同时进行内皮细胞和平滑肌细胞的培养,再分层种植于人造血管内腔,模拟正常血管壁结构。我们利用同一根脐带,同时获取内皮细胞和平滑肌细胞,方便有效、经济实用,以利于进一步进行相关研究。 1 材料和方法 1.1 人脐静脉内皮细胞原代培养 取一根长约30cm的脐带,于D-Hank’s液中4℃保存,6h内进行操作。先结扎两头的动脉,再将自制的两个冲洗接头分别插入脐静脉的两端,用D-Hank’s液冲净其内腔,再注入混合酶(0.01％Ⅰ型胶原酶3ml+0.25％胰蛋白酶15ml),夹闭后放于37℃消化20分钟。收集消化液,离心5
With the deepening of the study of artificial vascular endothelialization, endothelial cells and smooth muscle cells need to be cultured at the same time, and then stratified into artificial blood vessel lumen to simulate the normal vessel wall structure. We use the same umbilical cord, while obtaining endothelial cells and smooth muscle cells, convenient and effective, economical and practical, in order to facilitate further research. 1 Materials and methods 1.1 Human umbilical vein endothelial cell primary culture Take a cord about 30cm long, stored in D-Hank’s solution at 4 ℃, 6h operation. First ligation of the two arteries, and then two homemade flushing joints were inserted into the umbilical vein at both ends, with D-Hank’s liquid rinsed its lumen, and then mixed enzyme (0.01% collagenase type I 3ml + 0.25% trypsin 15ml), after cloning release at 37 ℃ for 20 minutes. Collect digestive juice and centrifuge 5