3D gene knockout tissue models using adult human stem cells, £90k

This project will develop and analyse GM/knockout adult stem cell lines in vitro, using novel, 3D self-assembling tissue-like environments that more closely mimic in vivo conditions. This tractable system will permit the analysis of the molecular events that control tissue development, organisation and function using human cells and uncomplicated by potential in vivo artefacts and species-dependent effects encountered in animal models. We will generate immortalised lines from primary human multipotent mesenchymal stem cells (MSCs) carrying genetic modifications, including stable gene knockout (rather than knockdown) MSC lines. These lines will be used in 3D mono- and multi-culture (representing bone and cartilage bi-differentiation) to focus on the mechanisms determining skeletal tissue differentiation and demonstrate the scientific strength of the proposal. The modular nature of this system allows sophisticated mechanistic analyses that are not possible using GM mice.