Cryopreservation of periosteum and whole bone by vitrification -establishment of organ freezing program.

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Cryopreservation of tissues and organs may play an important role in transplantation medicine. In the present study, chick bone marrow which was reamed from embryonic femur was cryopreserved with DMSO as cryo-protectant by slow rate (programmable biological freezer) or rapid (vitrification) cooling procedures, respectively. After thawing, they were implanted on the chorioallantoic membrane of a 9^<th> day chick embryo, and cultured for 10 days. The grafts were recovered and viability evaluated by soft X-ray photography (X-P) and histologically. The findings of X-Ps showed that the graft processed by slow rate cooling gave hypertropbic bone, whereas the mass was slightly inferior to that of the un-frozen control. On the other hand, the grafts cooled rapidly (vitrification) were atrophied. The histological findings suggested that the un-frozen control gave multi-layered periosteum and bone formation at the external periosteum and reamed cavity were observed. The graft after slow rate cooling also gave similar findings, but those after vitrification showed no bone formation. Overall results indicated that slow rate cooling with DMSO was suitable to keep post thaw viability of chick embryonic bone rather than cooling by vitrifiction.