摘要
在卵母细胞低温保存过程中,低温保护剂的添加与去除是必不可少的步骤。近年来微流控芯片被用于低温保护剂的添加及去除,而不同线型添加和去除低温保护剂会影响卵母细胞存活率及体外发育情况。利用以微流控芯片为中心的微混合系统,分别设计线性加载和去除保护剂的方案,并以分段线性衍生出凸型、凹型保护剂加载和去除方案,最终用上述方案共组合出9种添加-去除联用方案,研究不同线型添加-去除保护剂对卵母细胞存活率及体外发育情况的影响。结果表明:凹型添加-凸型去除联用方案下,猪MII期卵母细胞的存活率及桑椹胚率达到97.22%、46.03%,均显著优于其他添加-去除方法(P<0.05);低温保护剂添加及去除过程中,连续性添加及去除方案间具有一定的匹配关系,这将为优化微流控法连续性添加-去除低温保护剂方案提供新思路。
Cryoprotectants(CPA) addition and removal are essential steps in oocytes cryopreservation. In recent years, microfluidic chips have been used for addition and removal of CPA. The way of addition and removal in microfluidic chips will affect the survival rate and in vitro development of oocytes. In this paper, linear, convex and concave CPA loading or removal protocols were realized by adjusting injection models of microinjection pumps. Nine addition-removal combined protocols were achieved in the microfluidic system. The survival rate and in vitro development of oocytes with different CPA addition-removal protocols were investigated. Results showed that the survival rate(97.22%) and morula rate(46.03%) of porcine MII oocytes from concave loading-convex unloading protocol were significantly higher than that using other protocols(P < 0.05). There was a certain matching relationship between the addition and removal of CPA. This research provided a new solution for optimizing continuous addition-removal of CPA by microfluidic method.
作者
周新丽
杜羽琨
衣星越
戴建军
张德福
Zhou Xinli;Du Yukun;Yi Xingyue;Dai Jianjun;Zhang Defu(Institute of Biothermal Technology,University of Shanghai for Science and Technology,Shanghai 200093,China;Animal and Veterinary Research Institute,Shanghai Academy of Agricultural Sciences,Shanghai 201106,China)
出处
《中国生物医学工程学报》
CAS
CSCD
北大核心
2019年第6期719-725,共7页
Chinese Journal of Biomedical Engineering
基金
国家自然科学基金(51376132)
上海市自然科学基金(13ZR1429200)
