Accelerated formation of multicellular 3‐D structures by cell‐to‐cell cross‐linking

Abstract

The three-dimensional (3-D) arrangement of cells within tissues is integral to their development and function. Advances in stem cell science and regenerative medicine have stimulated interest in the replication of this architecture in vitro. We have developed a versatile method for controlling short-term cell-cell and cell-matrix interactions via a facile cell surface engineering process that enables the rapid formation of specific 3-D interactions for a range of cell types. We demonstrate that chemical modification of cell surfaces and matrix proteins can artificially accelerate the cell adhesion process and confirm the ability to control the formation of multicellular aggregates with defined architectures and heterotypic cell types. Direct comparison with a natural aggregation process seen during differentiation of embryonic stem (ES) cells revealed increased expression of developmental regulatory proteins and a concomitant enhancement of ES cell differentiation. Furthermore, this new methodology has numerous applications in generating layered structures. For example, we demonstrate improved transfer of therapeutic human keratinocytes onto a dermal layer in a skin repair model.

Controlled embryoid body formation via surface modification and avidin–biotin cross-linking

“…1a). Clearly engineering enhances cell-cell interaction, rapidly forming EBs in an avidin-dependent manner (De Bank et al 2007). Keeping scalability and cost effectiveness of the whole system in mind, 10 lg/mL avidin was chosen for all subsequent experiments.…”

“…We have previously described a novel 3D culture system for controlled aggregation of ES cells and enhanced EB formation (De Bank et al 2007). The method involves mild oxidation of surface sialic acid residues with sodium periodate to generate nonnative reactive aldehyde groups which are then biotinylated with biotin hydrazide (De Bank et al 2003).…”

Tissue engineering: strategies, stem cells and scaffolds

“…This process utilizes the affinity of Biotin to Avidin to provide a standardized and enriched system for differentiation ( Fig. 2) (De Bank et al 2007). This process has also been standardized by using cell suspensions, from which the cells are forced together using centrifugation (Burridge et al 2007).…”

Fluorescent oxygen sensitive microbead incorporation for measuring oxygen tension in cell aggregates

“…Application of the streptavidin-biotin interaction has been previously described for the assembly of multicellular aggregates [29,41,42]. Here, we further broaden the application range and report the efficacy of this conjugation method for integrating fluorescent probes into cartilaginous cell aggregates.…”