Department of Innovative Food Science, Mukogawa Women's University, Ikebiraki-cho 6-46, 663-8558 Nishinomiya, Japan.
Abi Inc., Ohtakanomori-higashi 1-12-1 270-0138, Nagareyama, Japan.
J Biochem. 2024 Mar 4;175(3):245-252. doi: 10.1093/jb/mvad087.
Regenerative medicine would benefit from a safe and efficient cryopreservation method to prevent the structural disruption caused by ice-crystal formation in cells and tissue. Various attempts have been made to overcome this problem, one of which is the use of an oscillating magnetic field (OMF). However, the underlying mechanism is unclear. In this study, to evaluate the effect of an OMF on ice-crystal formation in the leg muscles of mice, we used to use the frozen-section method with a slower freezing rate than is, usual which resulted in ice crystals forming in the tissue. We assessed the mean size and number per unit area of intracellular ice holes in sections of muscle tissue, with and without OMF. Ice-crystal growth was reduced in frozen tissue subjected to OMF. Furthermore, we evaluated the structure and function of proteins in frozen tissue subjected to OMF by immunostaining using an anti-dystrophin antibody and by enzymatic histochemistry for NADH-TR and myosin ATPase. The results imply that the ability of OMF to suppress ice-crystal growth might be related to their stabilization of bound water in biomolecules during freezing.
再生医学将受益于一种安全高效的冷冻保存方法，以防止冰晶形成对细胞和组织造成的结构破坏。人们已经尝试了各种方法来解决这个问题，其中之一是使用振荡磁场（OMF）。然而，其潜在机制尚不清楚。在这项研究中，为了评估 OMF 对小鼠腿部肌肉冰晶形成的影响，我们使用比通常更慢的冷冻速率的冷冻切片方法，导致组织中形成冰晶。我们评估了有和没有 OMF 的肌肉组织切片中细胞内冰晶孔的平均大小和单位面积数量。在受到 OMF 处理的冷冻组织中，冰晶的生长减少了。此外，我们通过使用抗肌营养不良蛋白抗体进行免疫染色和通过 NADH-TR 和肌球蛋白 ATP 酶的酶组织化学来评估受到 OMF 处理的冷冻组织中的蛋白质结构和功能。结果表明，OMF 抑制冰晶生长的能力可能与其在冷冻过程中稳定生物分子结合水的能力有关。
