Use of Tethered Hydrogel Microcoatings for Mesenchymal Stem Cell Equilibrium, Differentiation, and Self-Organization into Microtissues

Therapeutic adult mesenchymal stem cells (MSCs) lose multipotency and multilineage specialization in culture and after transplantation due to the absence of complex biological architecture. Here, it is shown that a transient ultrathin covering of permeable biomaterial can be differentially formulated to either preserve multipotency or induce multidifferentiation. Accordingly, populations of single, spherical MSCs in suspended media with high selec-tivity and specificity can be coated. Assembly of single, double, and triple hydrogel layers at MSC membranes is initiated by first attaching MSC-specific immunoglobulins onto CD90 or Stro-1 receptors and UEA-1 and soybean lec-tins. A secondary biotinylated immunoglobulin is targeted for avidin binding, which becomes an attractor for biotinylated alginate or hyaluronate, which are subsequently stiffened and gelled, in situ around the entire cell surface. Alginate microcoatings permeated with mobile BMP-2-induced osteospecial-ized tissue, vascular endothelial growth factor (VEGF) induced microcapillary formation, while microcoatings, with selected basement membrane proteins, preserve the multipotent phenotype of MSCs, for continuing rounds of cul-ture and directed specialization. Furthermore, forced packing of microcoated MSC populations creates prototypical tissue compartments: the coating par-tially simulating the extracellular matrix structures. Remarkably, microcoated MSC clusters show a tremendous simulation of a common embryological tissue transformation into the epithelium. Thus, confinement of free mor-phology exerts another control on tissue specialization and formation.