Reagents for Defining the Biological Environment of Three-Dimensional Cell Cultures

• Published in: Tissue engineering. Part A 2009-03, Vol.15 (3), p.713-713
• Main Authors: Angres, B, Benz, K, Sturany, S, Stoll, D, Wurst, H
• Format: Article
• Language: English
• ISSN: 1937-3341; 1937-335X

A novel system for the configuration of defined biological environments for three-dimensional cell culture was developed. This system consists of maleimide- and thiol-functionalized hydrophilic polymers, that can be modified with biofactors, such as adhesion molecules, growth factors, and extracellular matrix molecules and that subsequently can be cross-linked in the presence of cells to form a hydrogel. The chemistry allows uncomplicated setup of three-dimensional cell culture conditions in straightforward experimental steps. The selection of synthetic or natural polymers, such as polyvinyl alcohol and polyethylene glycol, or serum albumin, which elicit little or no cellular responses, provides an inert background for determining the effects of added specific biofactors. Different cell types, such as primary chondrocytes, mesenchymal stem cells, Madin-Darby Canine Kidney (MDCK) cells, NIH-3T3-cells, and Human Umbilical Vein Endothelial Cells (HUVEC) were successfully cultivated in different gel combinations. Furthermore, the effect of a fibronectin-derived adhesion peptide on the formation of cellular extensions in HUVEC cells is demonstrated and morphogenetic development of epithelial cell sheets in gels modified with extracellular matrix molecules is shown. Our approach provides convenient toolbox for establishing defined environments for 3-D cell culture and thus allows to determine cellular responses to extraneous factors as well as to study cell behavior under more controlled and adaptable conditions compared to traditional three-dimensional culture media. The hydrogel technology thus provides a versatile matrix that can be used to develop extracellular environments for tissue engineering and regenerative applications.