Combinatorial Development of Biomaterials for Clonal Growth of Human Pluripotent Stem Cells

Both human embryonic stem (hES) cells and induced pluripotent stem (hiPS) cells can self-renew indefinitely in culture, however current methods to clonally grow them are inefficient and poorly-defined for genetic manipulation and therapeutic purposes. Here we develop the first chemically-defined, xeno-free, feeder-free synthetic substrates to support robust self-renewal of fully-dissociated hES and hiPS cells. Materials properties including wettability, surface topography, surface chemistry and indentation elastic modulus of all polymeric substrates were quantified using high-throughput methods to develop structure/function relationships between materials properties and biological performance. These analyses show that optimal hES cell substrates are generated from monomers with high acrylate content, have a moderate wettability, and employ integrin α(v)β(3) and α(v)β(5) engagement with adsorbed vitronectin to promote colony formation. The structure/function methodology employed herein provides a general framework for the combinatorial development of synthetic substrates for stem cell culture.