活化小鼠卵母细胞的冷冻保存及解冻后合子重构

Cryopreservation of activated mouse oocytes and zygote reconstitution after thaw.

作者信息

Levron J, Willadsen S M, Shimmel T, Cohen J

机构信息

Gamete and Embryo Research Laboratory, Saint Barnabas Medical Center, NJ, USA.

出版信息

Hum Reprod. 1998 Dec;13 Suppl 4:109-16. doi: 10.1093/humrep/13.suppl_4.109.

DOI:10.1093/humrep/13.suppl_4.109

PMID:10091062

Abstract

A new approach to cryopreservation of unfertilized oocytes is proposed using techniques of artificial egg activation combined with nuclear transplantation. Matured mouse oocytes were released from metaphase II arrest by brief exposure to alcohol, allowed to progress to the pronuclear stage and then frozen according to a standard freezing protocol in propandiol. After thaw the female pronuclei were enucleated and fused with a male karyoplasts that were divided from in-vivo fertilized zygotes. Reconstituted zygotes, fresh and cryopreserved culture control zygotes were cultured to the blastocyst stage and transferred to pseudopregnant recipients. The rate of blastocyst formation was 75.8, 91.6 and 44.1% respectively. A total of 110, 215 and 70 blastocysts were transferred to pseudopregnant females respectively. The implantation rates were 36.4, 72.0 and 75.7% while the rates of fetal viability at mid-gestation were 15.5 (P < 0.0001), 51.1 and 37.1% respectively.

摘要

提出了一种结合人工卵母细胞激活技术与核移植技术的未受精卵母细胞冷冻保存新方法。将成熟的小鼠卵母细胞短暂暴露于酒精中使其从减数分裂中期II阻滞中释放出来，使其发育至原核期，然后按照标准冷冻方案在丙二醇中进行冷冻。解冻后，去除雌性原核，并将其与从体内受精的合子分离得到的雄性核质体融合。将重构合子、新鲜和合子冷冻保存的培养对照合子培养至囊胚期，并移植到假孕受体中。囊胚形成率分别为75.8%、91.6%和44.1%。分别将总共110、215和70个囊胚移植到假孕雌性动物体内。着床率分别为36.4%、72.0%和75.7%，而妊娠中期的胎儿存活率分别为15.5%（P<0.0001）、51.1%和37.1%。