玻璃冷凍造成cytoskeleton 受損包括: tubulin微管蛋白 & microtubules &microfilaments受損

卵子玻璃冷凍前使用 8 g/mL cytochalasin B 30min再冷凍可保護cytoskeleton, 提高胚胎分裂&囊胚率

Cryobiology. 2016 Mar 18. pii: S0011-2240(16)30016-5. doi: 10.1016/j.cryobiol.2016.03.005. [Epub ahead of print]

Stability of the cytoskeleton of matured buffalo oocytes pretreated with cytochalasin B prior to vitrification.

Wang CL1, Xu HY1, Xie L1, Lu YQ1, Yang XG1, Lu SS2, Lu KH3.

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Abstract

Stabilizing the cytoskeleton system during vitrification can improve the post-thaw survival and development of vitrified oocytes. The cytoskeleton stabilizer cytochalasin B (CB) has been used in cryopreservation to improve the developmental competence of vitrified oocytes. To assess the effect of pretreating matured buffalo oocytes with CB before vitrification, we applied 0, 4, 8, or 12 g/mL CB for 30 min. The optimum concentration of CB treatment (8 g/ml for 30 min) was then used to evaluate the distribution of microtubules and microfilaments, the expression of the cytoskeleton proteins actin and tubulin, and the developmental potential of matured oocytes that were vitrified-warmed by the Cryotop method. Western blotting demonstrated that vitrification significantly decreased tubulin expression, but that the decrease was attenuated for oocytes pretreated with 8 g/mL CB before vitrification. After warming and intracytoplasmic sperm injection, oocytes that were pretreated with 8 g/mL CB before vitrification yielded significantly higher 8-cell and blastocyst rates than those that were vitrified without CB pretreatment. The values for the vitrified groups in all experiments were significantly lower (P < 0.01) than those of the control groups. In conclusion, pretreatment with 8 g/mL CB for 30 min significantly improves the cytoskeletal structure, expression of tubulin, and development capacity of vitrified matured buffalo oocytes.