Human bone marrow mesenchymal stem cells (hBMMSCs) are capable of self-renewal and differentiating into multiple cell lineages, including osteoblasts, adipocytes, and chondrocytes. Osteoblasts are cells that form new bone with some osteoblasts turning into osteocytes during bone formation. This differentiation capacity holds great potential for cell therapies such as repair of bone defects. However, obtaining a sufficient number of cells to meet clinical demand is challenging due to limited sources, ageing of the cells and substantial variation in the bone regenerative capacity of cells from different donors and different isolation methods. Therefore, identification of markers capable of predicting the bone regenerative capacity of hBMMSCs is of great clinical importance. Alkaline phosphatase (ALP) is an enzyme that releases phosphate under alkaline conditions and is made in liver, bone, and other tissues. This enzyme is expressed at higher levels in hBMMSCs which undergo differentiation to osteoblasts cells. Therefore, there is great value in performing quantitative measurements for this important indicator, to enhance our understanding of the osteogenic differentiation process of hBMMSCs and to identify the most therapeutically potent donor cells for clinical applications. To this end, the aim of our project is to silence the expression of the ALP gene in hBMMSCs using gene therapy techniques and to assess their osteogenic potential compared to corresponding unmodified cells. Thereby we can determine if silencing of ALP expression reduces the capacity of these cells to form bone and can thereby act as a predictor of the bone regenerative potential of hBMMSCs.
