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

玻璃化冷冻保存骨膜和全骨-建立器官冷冻程序。

• 批准号: 14571407
• 负责人: TAKAHASHI Masanori
• 金额: $ 1.6万
• 依托单位: Tokyo Dental College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571407/
• 关键词: cryopreservation; periosteum; bone; DMSO; program freezing; vitrification; 急速凍結; 緩速凍結; 冷凍保存

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.

组织和器官的冷冻保存可能在移植医学中发挥重要作用。在本研究中，从胚胎股骨中取出的鸡骨髓分别通过慢速（可编程生物冷冻机）或快速（玻璃化）冷却程序以DMSO作为冷冻保护剂进行冷冻保存。解冻后，将它们植入第9天鸡胚的绒毛尿囊膜上，并培养10天。恢复移植物并通过软X射线摄影(X-P)和组织学评估活力。 X-Ps结果表明，经过慢速冷却处理的移植物产生了肥厚的骨，而质量略低于未冷冻的对照。另一方面，移植物迅速冷却（玻璃化）而萎缩。组织学结果表明，未冷冻对照具有多层骨膜，并且观察到外部骨膜和扩孔腔处的骨形成。缓慢冷却后的移植物也给出了类似的结果，但玻璃化后的移植物未显示出骨形成。总体结果表明，使用 DMSO 缓慢冷却比玻璃化冷却更适合保持鸡胚骨解冻后的活力。

项目成果

Takahasbi, M., Naniwa, T., Kaneko, S.: "Viability of the cryo-preserved chick embryonic bone by slow rate freezing and vitrification"Low temp.Med.. 29(3). 64-68 (2003)

Takahasbi, M.、Naniwa, T.、Kaneko, S.：“通过慢速冷冻和玻璃化冷冻保存的鸡胚骨的活力”Low temp.Med.. 29(3)。

高橋正憲, 浪花豊寿, 兼子悟: "プログラムフリーズ法およびガラス化法による幼若骨のviability温存の違い"Low Temp.Med.. 29.3. 64-68 (2003)

Masanori Takahashi、Toyotoshi Naniwa、Satoru Kaneko：“程序冷冻法和玻璃化法保存幼骨活力的差异”Low Temp.Med.. 29.3 (2003)。

高橋正憲, 浪花豊寿, 兼子悟: "プログラムフリーズ法およびガラス法による幼若骨のviability温存の違い"Low Temp.Med.. 29.3. 64-68 (2003)

Masanori Takahashi、Toyotoshi Naniwa、Satoru Kaneko：“使用程序冷冻法和玻璃法保存幼骨活力的差异”Low Temp.Med.. 29.3 (2003)。