生物体の凍結と細胞の生死を連結する階層的研究

Material type: 文書・図像類

Author: 林, 勇二郎ほか

Publisher: 金沢大学工学部

Publication date: 1999-03

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Note (General)：: 生物試料の凍結保存は,低温化と水分活性の低減により長期保存を図るものであるが,凍結・解凍の過程で様々な膠質的損傷が生じ,これが細胞の生残率を著しく低下させる原因となっている.細胞の生死に関わる膠質的損傷の究明には,マクロな伝熱過程と細胞の生死に関わる分子レベルの挙動を連結する試みが必須の課題となる....

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Material Type: 文書・図像類
Title: 生物体の凍結と細胞の生死を連結する階層的研究
Author/Editor: 林, 勇二郎
Hayashi, Yujiro
Author Heading: 林, 勇二郎
Hayashi, Yujiro
Publication, Distribution, etc.: 金沢大学工学部
Publication Date: 1999-03
Publication Date (W3CDTF): 1999-03
Alternative Title: Hierachical Study on Heat Transfer and Viability of Cell during Freezing of Biological Tissue
Periodical title: 平成10(1998)年度科学研究費補助金基盤研究(A) 研究成果報告書 = 1998 Fiscal Year Final Research Report
No. or year of volume/issue: 1995-1998
Volume: 1995-1998
Pages: 3p.-
Text Language Code: jpn
Subject Heading: 凍結保存
生物試料
生残率
凍結損傷の機序
ミクロ伝熱
Cryopresevation
Biological substance
Viability
Mechanism of freezing injury
Microscale Heat Transfer
Target Audience: 一般
Note (General): 生物試料の凍結保存は,低温化と水分活性の低減により長期保存を図るものであるが,凍結・解凍の過程で様々な膠質的損傷が生じ,これが細胞の生残率を著しく低下させる原因となっている.細胞の生死に関わる膠質的損傷の究明には,マクロな伝熱過程と細胞の生死に関わる分子レベルの挙動を連結する試みが必須の課題となる.本研究は,生物体の凍結の機序を,(1)マクロな伝熱過程,(2)細胞レベルでのミクロ挙動,さらには(3)分子レベルでの水の結合状態などの階層的な視点のもとで明らかにするとともに、それらに与えられた系統的な記述を連結させることにより,新しい輸送現象論を確立することを目的とする.前年度までに、(1)と(2)に対するミクロスコピックな速度論の展開と,(2)と(3)に対する実験的追究を進めてきた.最終年度である本年度は,(1)〜(3)の各階層に対する記述を連結させた輸送現象モデルの提示と計算を進めた.以下に得られた成果を記す.(1) 階層の中位にある細胞レベルの挙動,特に細胞内の氷晶形成を高い解像度で記述する速度論の追究を目的に,小麦ブロトプラストを供試したin vitroな凍結実験を行った.その結果,細胞外氷晶が膜表面での不均質核生成を発生させる要因であり,また,細胞膜の収縮はその作用を抑制することが明らかにされた.(2) 膜表面での不均質核生成(Surfec catalyzed nucleation:SCN)モデルに上記の動的因子を組み込んだ細胞凍結モデルが提示され、その有効性が検証された.(3) 各階層に対する記述を連結させ、マクロな伝熱過程を決定する操作パラメータ(冷却速度,凍害防御剤の濃度)ならびに生物試料(膜透過係数,組織体寸法)に対して凍結後の細胞の生残率を得るモデルが構築され,これによりマクロとミクロを連成した輸送現象論の確立が概ね達成された.
Freezing can slow down or stop some biological reactions for preserving the biological tissue, and thawing can bring it back to life. It is also true that freezing and thawing are lethal to the living system. During freezing, the extra- and intra-cellular ice formation, osmotic water permeation through cell membrane, deformation of cell and other behaviors occur at microscale and they bring serious injuries connecting to the life-and-death of living cells. These micro behaviors occurred on at least cell size level seem to be due to the physicochemical complexity of biological materials. Water is chemically restrained in the states of aqueous solution and colloidal solution, and those are physically confined in small spaces or compartmentalized by the cross-linked network structures.In this research project, firstly, simplified freezing model of cell element was proposed, and the extra- and intra-cellular ice formation, osmotic water permeation through cell membrane and other microscale behavior during the freezing of biological cell were discussed as a function of temperature.Secondly, viability of the cell was studied in relation with the micro-behaviors during freezing and thawing. In experiments, microscopic observation of wheat protoplasts was performed, and the cell viability was inspected by the fluorescence test. The osmotic conditions encountered during freeze-thaw process were also realized by osmotic manipulations. Summarizing these results, the cell damage during freezing process was estimated taking account of "solution effect injury" and "intracellular ice injury".Thirdly, the freezing process of biological tissue was numerically simulated using heat transfer and thermodynamic model. On the basis of these results, viability of the cell during freezing of biological tissue was discussed in conjunction with the heat transfer process.
研究課題/領域番号:07405011, 研究期間(年度):1995-1998
出典：「生物体の凍結と細胞の生死を連結する階層的研究」研究成果報告書　課題番号07405011 (KAKEN：科学研究費助成事業データベース（国立情報学研究所）)　　　本文データは著者版報告書より作成
DOI: 10.24517/00049274
Access Restrictions: 限定公開
Related Material: https://kaken.nii.ac.jp/search/?qm=30019765
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07405011/
https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-07405011/074050111998kenkyu_seika_hokoku_gaiyo/
Data Provider (Database): 国立情報学研究所 : 学術機関リポジトリデータベース（IRDB）（機関リポジトリ）
https://irdb.nii.ac.jp
Original Data Provider (Database): 金沢大学 : 金沢大学学術情報リポジトリKURA