Cryopreservation of antifreeze protein multicellular systems based on X-ray nanoscale observations in vivo for ice crystals

2022 Fiscal Year Final Research Report

• Project/Area Number: 21K14466
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 27040:Biofunction and bioprocess engineering-related
• Research Institution: Ibaraki University
• Principal Investigator: Kuramochi Masahiro 茨城大学, 理工学研究科(工学野), 助教 (60805810)
• Project Period (FY): 2021-04-01 – 2023-03-31
• Keywords: 不凍タンパク質 / 線虫 / 凍結保存 / 氷晶制御 / 熱ヒステリシス / 時分割X線計測

Outline of Final Research Achievements

In the current study, I aimed to establish the most effective cryopreservation method for AFP. I verified AFP effectiveness and analysed the AFP-ice interaction by X-ray nanoscale measurement in vivo. I found that the survival rate of transgenic C. elegans expressing AFP at body wall muscles was significantly higher than that of the wld-type control animals. Next, X-ray nanoscale measurements were applied to monitor AFP-induced ice crystal regulation in vivo. Interestingly, I found that the molecular dynamics of the wild-type AFP and the loss-of-function AFP mutant behaved qualitatively differently at -5 and -10°C. These results indicate that the inhibition of ice crystal growth by AFP also acts as an important function in cryopreservation in vivo.

Free Research Field

生物物理

Academic Significance and Societal Importance of the Research Achievements

不凍タンパク質(Antifreeze Protein: AFP)は、氷晶成長を抑制するユニークな機能をもつ。本研究から、多細胞生物である線虫の凍結保存において、AFPが有効に働くことがわかった。その分子メカニズムとして、氷晶成長抑制という機能が動物体内でも発揮している様子をX線ナノスケール計測によって捉えることができた。これらの知見は、細胞や組織、臓器に至るさまざまな生体材料の凍結保存において、AFPが効果的に機能する可能性を示している。将来的に、臓器保存などを課題とする医療・医学分野での技術展開が期待できる。