Abstract
The inhibition of unspecific adhesion of human white blood cells is a prerequisite for applications requiring the control of defined surface interactions. In this study, a passivation agent based on polyethylene glycol (PEG) for glass surfaces was investigated for the use with human peripheral blood mononuclear cells (PBMC). The grafting of 2000 g/mol methoxy-terminated PEG-urea-triethoxysilane (mPEG2000) onto glass surfaces successfully inhibited unspecific spreading of both human PBMC and platelets in all experiments. The prevention of surface interactions was independent on the anticoagulant used during blood collection. The total efficiency to prevent even transient immobilization of PBMC to the PEG modified surfaces was 97 ± 2%. This makes the passivation with PEG a well suited surface modification for preventing unspecific surface interaction in order to study only defined surface interactions of human PBMC.Electronic supplementary materialThe online version of this article (doi:10.1186/1559-4106-8-14) contains supplementary material, which is available to authorized users.
Highlights
A striking difference between in vivo and in vitro systems is the adhesion signalling
A passivation agent based on polyethylene glycol (PEG) for glass surfaces was investigated for the use with human peripheral blood mononuclear cells (PBMC)
In order to investigate the potential of a PEG-layer to prevent surface interactions of PBMC, PEGylated glass cover slips were prepared
Summary
A striking difference between in vivo and in vitro systems is the adhesion signalling. To date, isolated cells for ex-vivo studies are usually cultured on standardized plastic cell culture dishes, both in immunology and other fields of biology. These dishes are commonly based on polystyrene, with a one type of PBMC, are characterized by a distinct adhesion to a variety of different materials and molecules. Integrin αMβ2 (CD11b/CD18 or Mac-1) and αXβ2 (CD11c/CD18) bind to surfacebound C3b, both on PEG-like surfaces [11] and other biomaterials [12] This complement binding might even in serum- and plasma-free conditions aid the adhesion as monocytes are capable of synthesising complement [13]. We aimed to investigate the effectiveness of a PEG-mediated passivation in order to prevent all surface interactions of PBMC in serum-free conditions by use of PEGmonolayers grafted to glass cover slips
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