Detection of living cells in non-processed but deep-frozen bone allografts

Impacted morselized donor bone is successfully used to treat bone loss in revision total hip arthroplasties. It is generally thought, but not proven, that the processing and storage at -80 degrees C of the donor bone kills all cells. Because of the risk of contamination and to increase our understanding about the process of new bone formation after revision total hip arthroplasty, the aim of this study was to investigate whether the donor bone does contain vital cells. Samples from 11 femoral heads were obtained according to the American and European standards of bone banking, and tested for their capacity to give rise to proliferating cells, using tissue culture methods. All bone samples were stored at - 80 degrees C for a minimum of 6 months. Bone sample cores were morselized and cultured for 6 weeks. Inverted phase contrast microscopy was used to evaluate cell growth. DNA marker analysis was used to confirm cellular identity. All bank bone samples gave rise to cell growth. The cell cultures showed osteoblastic characteristics in that they expressed high levels of alkaline phosphatase activity. DNA marker analysis showed identical alleles for cultured cells from frozen bone and freshly obtained buccal cells from the same donor, indicating that the cells growing from the banked bone were indeed originating from the donor tissue. It was therefore concluded that -80 degrees C freezing of bone tissue does not routinely kill cells within the tissue.