Stem cell plasticity, osteogenic differentiation and the third dimension

Different cues present in the cellular environment control basic biological processes. A previously established 3D microwell array was used to study dimensionality-related effects on osteogenic differentiation and plasticity of marrow stromal cells. To enable long-term culture of single cells in the array a novel surface functionalization technique was developed, using the principle of subtractive micro contact printing of fibronectin and surface passivation with a triblock-copolymer. Immunohistochemical stainings showed that when cultivated in 3D microenvironments, marrow stromal cells can be maintained in the wells for up to 7days and be induced to commit to the osteogenic lineage. In conclusion, this work shows the modification of a 3D microwell array allowing the long term study of single stem cell plasticity and fate in controlled microenvironment