Modeling Osteosarcoma Using Li-Fraumeni Syndrome Patient-derived Induced Pluripotent Stem Cells

This Li-Fraumeni syndrome iPSC-based osteosarcoma model can help answer key questions in the osteosarcoma research field about using patient iP cells in cancer modeling. The main advantage of this model is that it allows the generation unlimited cells at all stages of osteosarcoma development for mechanistic studies, biomarker identification, and drug screening. Ruoji will be demonstrating the cell culture procedure and I will be demonstrating the animal procedure.

After two weeks of mouse embryonic fibroblast, or MEF, co-culture wash the 80 to 90%confluent iPS cell culture with five milliliters of DPBS followed by the addition of one milliliter of cell detachment solution to the cells for a three minute incubation at room temperature. It is critical to maintain iPS cells in good conditions on MEFs for at least 14 days for the cells to be able to attach to a gelatin-coated plate for their differentiation into mesenchymal stem cells. When the cells have detached from the plate bottom add nine milliliters of MEF/MSC culture medium to the cells to neutralize the cell detachment solution and transfer the cell suspension into an uncoated 100 millimeter tissue culture plate.