Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, SK, Canada.
Agriculture and Agri-Food Canada, Saskatoon Research and Development Center, Canadian Animal Genetic Resource Program, Saskatoon, SK, Canada.
PLoS One. 2020 Sep 2;15(9):e0238573. doi: 10.1371/journal.pone.0238573. eCollection 2020.
Sugars are commonly supplemented into vitrification solution to dehydrate cells in order to reduce the formation of fatal intracellular ice crystals. Natural honey is a mixture of 25 sugars (mainly fructose and glucose) that have different biological and pharmacological benefits. The present study was designed to determine if honey can be used as a nonpermeating cryoprotectant in vitrification of bovine oocytes. In the first experiment, denuded-MII oocytes were exposed to 0.25, 0.5, 1.0, 1.5 or 2.0 M of honey or sucrose. Natural honey and sucrose caused similar ooplasm dehydration. A significant relationship existed between time and ooplasm volume change (P < 0.05), during dehydration and rehydration phases, in both honey and sucrose solutions. In the second experiment, the immature cumulus-oocyte complexes (COCs) were vitrified in an EG/DMSO-based vitrification solution containing honey (0.5, 1 or 1.5 M) or sucrose (0.5 M) as a gold standard. The vitrified-warmed COCs were matured in vitro and evaluated for nuclear maturation. The maturation (MII) rate was greater in nonvitrified control (81%) than vitrified groups (54%, P < 0.05). In the third experiment, COCs were either remained nonvitrified (control) or vitrified in 1.0 M honey or 0.5 M sucrose, followed by IVM, IVF and IVC (for 9 days). Cleavage rate was greater in control (74%) than in vitrified groups (47%, P < 0.05), without significant difference between sugars. Blastocyst rate was 34, 13 and 3% in control, honey and sucrose groups respectively (P < 0.05). In conclusion, natural honey acted as a nonpermeating cryoprotectant in vitrification solution and improved the embryonic development in vitrified bovine COCs.
糖通常被添加到玻璃化溶液中,以使细胞脱水,以减少致命的细胞内冰晶的形成。天然蜂蜜是 25 种糖(主要是果糖和葡萄糖)的混合物,具有不同的生物学和药理学益处。本研究旨在确定蜂蜜是否可用作牛卵母细胞玻璃化的非渗透型冷冻保护剂。在第一个实验中,裸 MII 卵母细胞暴露于 0.25、0.5、1.0、1.5 或 2.0 M 的蜂蜜或蔗糖中。天然蜂蜜和蔗糖导致相似的卵浆脱水。在脱水和复水阶段,蜂蜜和蔗糖溶液中的卵浆体积变化与时间之间存在显著关系(P < 0.05)。在第二个实验中,不成熟的卵丘-卵母细胞复合物(COC)在含有蜂蜜(0.5、1 或 1.5 M)或蔗糖(0.5 M)的基于 EG/DMSO 的玻璃化溶液中进行玻璃化。玻璃化-加热的 COC 在体外成熟,并评估核成熟。非玻璃化对照组(81%)的成熟率(MII)大于玻璃化组(54%,P < 0.05)。在第三个实验中,COC 保持非玻璃化(对照)或在 1.0 M 蜂蜜或 0.5 M 蔗糖中玻璃化,然后进行 IVM、IVF 和 IVC(持续 9 天)。对照组的卵裂率(74%)大于玻璃化组(47%,P < 0.05),但糖之间没有差异。对照组、蜂蜜组和蔗糖组的囊胚率分别为 34%、13%和 3%(P < 0.05)。总之,天然蜂蜜在玻璃化溶液中作为非渗透型冷冻保护剂,提高了玻璃化牛 COC 的胚胎发育。