Effect of Magnetized Freezing Extender on Membrane Damages, Motility, and Fertility of Boar Sperm Following Cryopreservation

Abstract

Magnetized water is defined as the amount of water that has passed through a magnet. The magnetic field weakens the hydrogen bonds between the water molecules, leading to the magnetized liquid acquiring special characteristics such as easy supercooling and forming smaller ice crystals. We researched the influences of a magnetized freezing extender on cell membrane damage and in vitro fertilization of boar sperm during cryopreservation. The freezing extenders were passed through 0, 2000, 4000, and 6000 gausses (G) of magnetic devices using a liquid cycling pump system and then used for the sperm freezing process. The damage to plasma, acrosomal, and mitochondrial membranes in frozen-thawed spermatozoa was investigated by flow cytometry, and motility was assessed using the CASA system. The fertility of frozen-thawed sperm was estimated using in vitro fertilization. The damage to the membranes was significantly decreased in the magnetized freezing extender by the 6000 G magnetic field compared to that of the control in frozen-thawed sperm, and motility was increased in the 6000 G group. Although there were no significant differences in the cleavage rates of in vitro fertilized oocytes among the treatment groups, the ratio of blastocyst formation increased in the magnetized freezing extender groups compared with that in the control group. The number of blastocysts was significantly higher in the 4000 G group than in the 0 G group. In conclusion, these results suggest that a magnetized freezing extender could improve the freezability of sperm and the development of oocytes fertilized in vitro with frozen-thawed sperm.

Incorporate delivery, warming and washing methods into efficient cryopreservation

“…The devices for vitrification are various, such as cyrotop (for oocytes) ( Miao et al, 2022 ), plastic straw (for spermatids) ( Patra et al, 2021 ), and cryomesh (for islets) ( Zhan, Rao, et al, 2022 ). However, low temperature can cause a range of damage to biological samples, including protein denaturation ( Chen et al, 2022 ), membrane damage ( Lee et al, 2023 ), oxidative stress ( Gualtieri et al, 2021 ). Since DMSO was first used in bull sperm cryopreservation in 1959, it has been found that the addition of a certain concentration of DMSO could resist these cryodamages ( Lovelock and Bishop, 1959 ; Stubbs et al, 2020 ).…”