[Mechanical brush-catheter abrasion method for the isolation and culture of human umbilical vein endothelial cells. First in vitro results].
Academic Article
Overview
abstract
UNLABELLED: Mechanical brush-catheter abrasion method for the isolation and culture of human umbilical vein endothelial cells. First in vitro results. PURPOSE: 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). MATERIAL AND METHODS: 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. RESULTS: 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. CONCLUSIONS: The brush-catheter abrasion is an effective method for collecting viable human endothelial cells in vitro.
