Effects of partially replacing glycerol with cholesterol-loaded cyclodextrin on protamine deficiency, in vitro capacitation and fertilization ability of frozen-thawed Yanbian Yellow cattle sperm.

Glycerol is widely used as a cryoprotectant to protect the sperm from freezing damage during cryopreservation. However, glycerol at a high concentration has toxic effects on the sperm. Therefore, we explored the effects of partially replacing glycerol with cholesterol-loaded cyclodextrin (CLC) in a cryoprotectant on protamine deficiency, in vitro capacitation, and fertilization ability of freeze-thawed Yanbian Yellow cattle sperm. We used fresh semen, control (6% glycerol), and four treatment-I, II, III, and IV (3% glycerol + 0, 0.75, 1.5, and 3 mg/mL CLC, respectively)-groups. Computer-assisted semen analysis; JC-1, CMA3, and FluoZin-3-AM staining; flow cytometry; and IVF were conducted. Replacing a portion of glycerol with 1.5 mg/mL CLC significantly improved sperm motility, viability, plasma membrane integrity, acrosome integrity, and membrane lipid disorders, mitochondrial membrane potential (MMP), capacitation, and fertilization ability (P < 0.05) compared with the control. Additionally, in group I and III, the protamine deficiency were significantly lower (P < 0.05) than in the control group. It was found that 6% glycerol has a higher degree of damage to sperm DNA integrity than 3% glycerol. Overall, this study revealed that partial replacement of glycerol with CLC can be used as a novel cryoprotection method to reduce the toxicity of glycerol and improve the quality of thawed Yanbian Yellow cattle sperm.