Principle study of developing a tissue preservation bank for pathological specimen

病理标本组织保存库开发原理研究

• 批准号: 14570139
• 负责人: TAKAHASHI Rei
• 金额: $ 1.73万
• 依托单位: KYOTO UNIVERSITY
• 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-14570139/
• 关键词: Tissue preservation; Freeze-drying; Genetic analysis; Tissue bank; Ice crystal; Biological macromolecule; Vitrification; 組織保存; トレハロース; ガラス化状態

Because of the routine needs of our handling pathological specimens, we have developed a protocol that allows appropriate methods of preserving specimens suitable for molecular and genetic analyses. Using the principles of freeze-drying, pathological tissues are found to be stably preserved at room temperature for a long period of time. In this project, we have elucidated the following points based of the experiments performed.1)Macromolecules including protein, DNA and RNA can be stably stored after years of preservation under the conditions we have obtained. Addition of trehalose or EDTA in the freeze-dried specimen favorably inhibits RNA degradation in the course of preservation.2)Macromolecules can well be preserved if oxygen and moisture are avoided during the preservation period. Temperature is not a critical factor, therefore, freeze-dried specimens can be stored at room temperature instead of being kept in a deep freezer.3)The freeze-substitution protocol instead of freeze-drying also gives similar outcome in preserving DNA and RNA if the protein analysis is not necessary.4)Cytological smear specimens are applicable for our protocol.

由于我们处理病理标本的常规需要，我们开发了一种方案，允许采用适当的方法保存适合于分子和遗传分析的标本。利用冷冻干燥原理，病理组织可以在室温下稳定保存很长一段时间。在本项目中，我们在实验的基础上阐明了以下几点：1)在所获得的条件下，包括蛋白质、DNA和RNA在内的大分子在多年保存后可以稳定地保存下来。在冻干样品中添加海藻糖或EDTA有利于抑制RNA在保存过程中的降解。2)如果在保存期间避免氧气和水分，大分子可以得到很好的保存。温度不是一个关键因素，因此，冷冻干燥的标本可以在室温下保存，而不是保存在深冻冰箱中。3)如果不需要进行蛋白质分析，冷冻替代方案在保存DNA和RNA方面也可以得到类似的结果。4)细胞学涂片标本适用于我们的方案。

项目成果

Yazumki, S. et al.: "Intraductal ultrasonography aids detection of carcinoma in situ in a patient with a choledochal cyst"Gastrointestinal Endoscopy. (in press). (2003)

Yazumki, S. 等人：“导管内超声检查有助于检测胆总管囊肿患者的原位癌”胃肠内窥镜检查。

T.Matsushita et al.: "Suppressed complement activation in human Decay accelerating factor transgenic procine Liver cross-circulated with nonhuman Primates."Transplantation. 75:11. 1807-1812 (2003)

T.Matsushita 等人：“人衰变加速因子转基因猪肝脏与非人灵长类动物交​​叉循环中的补体激活被抑制。”移植。

Kano, Y. et al.: "Immunosuppressant FK5O6 induces neurite outgrowth in PC12 mutant cells with impaired NGF-promoted neuritogenesis via a novel MAP kinase signaling pathway"Neurochem. Res.. 27・12. 1655-1661 (2002)

Kano, Y. 等人：“免疫抑制剂 FK5O6 通过新的 MAP 激酶信号通路诱导 NGF 促进的神经突发生受损的 PC12 突变细胞中的神经突生长”Neurochem. 1655-1661 (2002)。

T.Okuyama et al.: "Detection of trichomonas vaginalis by combinded A6p and P65 sequences using PCR amplification From papanicolaou-stained smears."ACTA Cytologica. 47:4. 696-698 (2003)

T.Okuyama 等人：“使用来自巴氏染色涂片的 PCR 扩增，通过组合 A6p 和 P65 序列检测阴道毛滴虫。”ACTA Cytologica。

J.Oh et al.: "Mutation in two matrix metalloproteinase Gene, MMP-2 and MT1-MMP, are synthetic Lethal in mice."Oncogene. (in press).

J.Oh 等人：“两种基质金属蛋白酶基因 MMP-2 和 MT1-MMP 的突变对小鼠来说是合成致死的。”癌基因。