Abstract

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.

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