在没有程序降温仪辅助的情况下冷冻哺乳动物胚胎。

Freezing of mammalian embryos without the aid of a programmable freezer.

作者信息

Boone W R, Brown C A, Vasquez J M, Shapiro S S

机构信息

Department of Obstetrics and Gynecology, University of Wisconsin, Madison 53792.

出版信息

Fertil Steril. 1988 Aug;50(2):348-54. doi: 10.1016/s0015-0282(16)60085-7.

DOI:10.1016/s0015-0282(16)60085-7

PMID:3396704

Abstract

A simple, inexpensive and repeatable method of freezing/thawing (F/T) mammalian concepti was developed with the use of 2-cell mouse embryos. Cryoprotectants, length of exposure to protectants, and subzero holding times before liquid nitrogen (LN2) exposure were examined in an effort to obtain an effective freezing protocol. Sequential examination of these variables provided data suggesting that 3.5 M dimethyl sulfoxide (DMSO) plus 0.25 M sucrose exposure for 5 minutes at room temperature, followed by a -30 degrees C environment for 90 minutes, best prepared embryos for LN2 storage. After thawing and culture, 48 of the 93 (52%) embryos developed to the blastocyst stage.

摘要

利用2细胞期小鼠胚胎，开发了一种简单、廉价且可重复的哺乳动物胚胎冻融（F/T）方法。为了获得有效的冷冻方案，研究了冷冻保护剂、接触保护剂的时间长度以及暴露于液氮（LN2）之前的零下保存时间。对这些变量的顺序检查提供的数据表明，在室温下用3.5 M二甲基亚砜（DMSO）加0.25 M蔗糖处理5分钟，然后在-30℃环境下放置90分钟，能使胚胎为LN2保存做好最佳准备。解冻和培养后，93个胚胎中有48个（52%）发育到囊胚阶段。