Biochemical isolation and identification of microvascular endothelial cells

ABSTRACT

To overcome the problems commonly involved in the culture of microvascular endothelial cells included unreliable isolation techniques and low cell-yields, a simplified protocol for the selective cultivation of rat heart microvascular endothelial cells [RHMEC], based on the cell specific expression of platelet-endothelial cell adhesion molecule PECAM-1 [CD31], was developed. Subconfluent primary cultures consisted of a mixture of endothelial cells, fibroblasts, and smooth muscle cells were isolated by their selective binding to magnetic beads coupled with anti-PECAM-I monoclonal antibody. After 2 immunomagnetic purification steps, a homogenous population of RHMEC was obtained, which showed typical cobblestone morphology, expressed CD3 1 and Von Willebr and factor proliferated in response to endothelial growth supplement and formed capillary-like tubes in a 3-dimensional ESH-gel matrix. This simple technique might facilitate a more widespread use of microvascular endothelial cell cultures obtained from different human or animal organs for functional in vitro studies