Mild Quantitative One Step Removal of Macrophages from Cocultures with Human Umbilical Vein Endothelial Cells Using Thermoresponsive Poly(Di(Ethylene Glycol)Methyl Ether Methacrylate) Brushes.

We report on a mild, label-free and fast method for the separation of human umbilical vein endothelial cells (HUVEC), which are relevant cells, whose use is not limited to studies of endothelial dysfunction, from co-cultures with macrophages to afford HUVEC in ∼100% purity. Poly(di(ethylene glycol)methyl ether methacrylate) (PDEGMA) brushes with a dry thickness of (5 ± 1) nm afford the highly effective one step separation by selective HUVEC detachment, which is based on the brushes' thermoresponsive behavior. Below the thermal transition at 32°C the brushes swell and desorb attached proteins, resulting in markedly decreased cell adhesion. Specifically, HUVEC and macrophages, which were differentiated from THP-1 monocytes, were seeded and attached on PDEGMA brushes at 37°C. After decreasing the temperature to 22°C, HUVEC showed a decrease in their cell area, while the macrophages were not markedly affected by the temperature change. After mild flushing with cell culture medium, the HUVEC could be released from the surface and reseeded again with ∼ 100% purity on a new surface. With this selective cell separation and removal method, it was possible to separate and thereby purify HUVEC from macrophages without the use of any releasing reagent or expensive labels, such as antibodies. This article is protected by copyright. All rights reserved.