An effective method of isolating microvascular endothelial cells from the human dermis

• DOI: 10.1002/cbin.11448
• Publication Date: 2020-08-27T19:29:33Z
• Authors: Hui Hou; Jiao Li; Ling Zhou; Jiannan Liang; Juanjuan Wang; Junqin Li; Ruixia Hou; Juan Li; Xiaohong Yang; Kaiming Zhang
• Keywords: Percoll; Foreskin; CD31

Abstract Dermal microvascular endothelial cells (DMECs) play central roles in inflammation and angiogenesis have become important cell models for studying various skin diseases. However, primary DMECs are difficult to culture often contaminated by mesenchymal stem cells, fibroblasts, other stromal cells. Surgically removed superfluous foreskin was first cut into pieces, digested with two types of enzymes, dispersed single Cells obtained from the dermis were then subjected Percoll density gradient centrifugation located between densities 1.033 1.047 g/ml further purified growth medium containing decreasing concentrations puromycin. Obtained HDMECs identified microscopy, flow cytometry, quantitative reverse‐transcription polymerase chain reaction, western blot analysis, immunofluorescent staining. The expression CD31 (PECAM‐1), CD34, VEGFR2, VWF (Von Willebrand Factor), VE‐Cadherin (CD144), NOS positive. found abilities uptake acetylated low‐density lipoprotein. Growth curves viability analyzed, a pattern consisting “latency phase‐logarithmic phase‐stagnation phase” determined. In this study, simple, rapid, effective, low‐cost method is established isolate purity over 91% high viability. showed good repeatability allowed stable passage. This study provides technical support theoretical guidance physiological characteristics HDMECs, pathogenesis associated,