Materials and methods for preparing mesenchymal stem cells are described such that the mesenchymal stem cells remain for a longer period of time than would normally be found at a site of tissue healing. The mesenchymal stem cells are pretreated to exhibit osteogenic characteristics and, when combined with extracellular matrix components produced from the pretreated cells, improve tissue healing. Co-administration of the pretreated cells and their secreted extracellular matrix components at an injured site or defect allowed the mesenchymal stem cells to co-localize at or near the injured or defect site for a longer period. The combination improves overall repair of the tissue by extending the retention signal of mesenchymal stem cells. A three-dimensional scaffold containing the pretreated cells with or without addition of the secreted extracellular matrix is also described. The conditioned scaffold is suitable for use in vivo and may be preserved cryogenically prior to use.
