Non-unions are among the most severe complications in trauma surgery and orthopedics. Functional impairment and persistent pain substantially reduce patients’ quality of life. In the treatment of large bone defects and non-unions, autologous cancellous bone grafting remains the method of choice due to its osteogenic, osteoinductive, and osteoconductive properties. However, failure rates of more than 40% have been reported. One potential contributing factor is the ischemia time between graft harvesting and transplantation: The preliminary data demonstrate that as little as 30 minutes of ischemia significantly alters the cellular stress response. Cold preservation or storage in histidine-tryptophan-ketoglutarate (HTK) solution may attenuate these effects. The aim of this study is to investigate the effects of ischemia time, cold preservation, and HTK storage on the viability and function of murine cancellous bone grafts both in vitro and in vivo in appropriate mouse models. Ultimately, this study aims to optimize the intraoperative management of bone grafts and improve bone healing rates.