Isolation and pure culture of microvascular endothelial cells from the fetal skin.

Abstract

Microvascular endothelial cells were purely isolated from human fetal skin using magnetic particles. The principle of this technique is based on the selective binding of the lectin Ulex europaeus I (UEA I) to the endothelial cell surface via fucose residues. Initially UEA I was covalently bound to tosyl-activated magnetic polydisperse polymer particles (Dynabeads) and then the UEA I-coated beads were collected using a magnetic particle concentrator (MPC). Endothelial cells were isolated by extracting microvascular segments from trypsin-treated fetal skin tissue and were purified by sieving with nylon mesh and by 35% Percoll gradient centrifugation. For further purification, the obtained cells were incubated with UEA I-coated Dynabeads. The endothelial cells bound to the Dynabeads were collected using MPC. This is a simple and reproducible technique for isolating a pure population of microvascular endothelium from the fetal skin.