Priebe M, Paulsen F, Jahnke T, Grimm J, Heller M, Müller-Hülsbeck S
Klinik für Diagnostische Radiologie, Institut für Anatomie, Kiel, Germany.
Rofo. 2001 Oct;173(10):955-8. doi: 10.1055/s-2001-17586.
Mechanical brush-catheter abrasion method for the isolation and culture of human umbilical vein endothelial cells. First in vitro results.
To determine the value of a nylon brush (Cragg thrombolytic catheter system) with regard to the isolation and culture of viable human vascular umbilical vein endothelial cells (HUVEC).
Fresh human umbilical cords were collected after a maximum of 6 - 8 h post delivery. They were stored in sterile tubes till further preparation. After predilation the umbilical cord veins, a 6 French brush catheter was inserted, expanded, and removed by hand using rotating movements. The nylon brush was shaken out in cell medium containing serum proteins by a connected motor drive. After repeating these procedures, the detached endothelial cells were centrifugated and resuspended. The cell numbers were assessed and cell viability controlled.
The endothelial cell (EC) viability depended on the umbilical cord's age and was the highest (29.8 +/- 11.4 %) within 3 h after delivery. The brush abrasion (3.32 x 10(5) +/- 52016 viable EC/10 cm vein segment) enabled us to culture these cells under different cell concentrations. They grew to confluence after 2 - 3 weeks.
The brush-catheter abrasion is an effective method for collecting viable human endothelial cells in vitro.
用于人脐静脉内皮细胞分离和培养的机械刷式导管磨损法。首次体外实验结果
确定尼龙刷（Cragg溶栓导管系统）在分离和培养有活力的人血管脐静脉内皮细胞（HUVEC）方面的价值。
在分娩后最多6 - 8小时收集新鲜人脐带。将它们储存在无菌管中直至进一步制备。在对脐带静脉进行预扩张后，插入一根6F刷式导管，扩张并通过手动旋转动作取出。通过连接的电机驱动在含有血清蛋白的细胞培养基中抖出尼龙刷。重复这些步骤后，将分离的内皮细胞离心并重悬。评估细胞数量并控制细胞活力。
内皮细胞（EC）活力取决于脐带的年龄，在分娩后3小时内最高（29.8 +/- 11.4%）。刷式磨损（3.32 x 10(5) +/- 52016个有活力的EC/10 cm静脉段）使我们能够在不同细胞浓度下培养这些细胞。它们在2 - 3周后生长至汇合。
刷式导管磨损是一种在体外收集有活力的人内皮细胞的有效方法。
