微流体装置线性去除猪MII期卵母细胞冷冻保护剂的实验研究

发布时间：2019-07-02 15:36

【摘要】：低温保存后的卵母细胞在使用前必须要去除冷冻保护剂,目前常用的分步法去除步骤繁琐,容易丢失细胞,而且会对细胞造成致命的渗透损伤。为减小细胞渗透损伤,设计制作适合卵母细胞保护剂去除的微流体装置,研究微流控线性法去除猪MII期卵母细胞低温保护剂时在不同时间(6、8、10 min)下卵母细胞的体积变化,以及对卵母细胞存活率与发育率的影响;并与传统的去除方法(一步法和分步法)进行比较。结果表明,采用微流体装置线性去除冷冻保护剂,8 min为实验中的最优去除时间;线性法能够明显减小细胞的渗透损伤,其最大归一化渗透膨胀体积为1.12±0.07,卵母细胞的存活率、卵裂率及囊胚率分别达到83.6%、72.4%、21.5%,均显著高于一步法和分步法(P0.05)。因此,微流控线性去除冷冻保护剂能够显著减小细胞的渗透损伤,为卵母细胞低温保存技术提供新思路。
[Abstract]:Cryopreserved oocytes must remove cryoprotectants before using them. At present, the common step-by-step removal steps are tedious, easy to lose cells, and can cause fatal osmotic damage to cells. In order to reduce cell osmotic injury, a microfluidic device suitable for oocyte protectant removal was designed and made to study the changes of oocyte volume at different time (6, 8, 10 min) and the effect on oocyte survival and development rate at different time (6, 8, 10 min) when the cryopreservation agent was removed by microfluidic linear method, and compared with the traditional removal methods (one-step method and step-by-step method). The results showed that 8 min was the optimal removal time in the experiment, and the maximum normalized osmotic expansion volume was 1.12 卤0.07, and the survival rate, blastocyst rate and blastocyst rate of oocytes were 83.6%, 72.4% and 21.5%, respectively, which were significantly higher than those of one-step method and step-by-step method (P 0.05). Therefore, microfluidic linear removal of cryoprotectants can significantly reduce the osmotic damage of cells and provide a new idea for cryopreservation of oocytes.
【作者单位】：上海理工大学生物热科学研究所;上海市农业科学院畜牧兽医研究所;
【基金】：国家自然科学基金(51376132)
【分类号】：R318.52

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