Isolation, Culture and Identification of Human Microvascular Endothelial Cells: A Comparison of Abdominal Subcutaneous and Omental Adipose Tissue

Abstract

The isolation and characterisation of human microvascular endothelial cells from adipose tissue is described and a comparison is made between isolations from abdominal subcutaneous and omental adipose tissue. Microvascular cells were isolated by incubating finely chopped material in (i) collagenase type II solution, followed by density gradient centrifu-gation and further incubation in trypsin, or in (ii) collagenase type II solution, followed by trypsin incubation and selective filtration. The final step in both procedures was selection using Ulex europaeus agglutinin-1 coated Dynabeads. The cells were characterised using immunocytochemical and biochemical techniques. The cells exhibited characteristic staining for antigens recognised by antibodies against von Willebrand factor and platelet-endothelial cell adhesion molecule (PECAM-I). They also expressed angiotensin-converting enzyme and internalised acetylated low-density lipoprotein.