Kasai M
College of Agriculture, Kochi University, Japan.
Mol Biotechnol. 1997 Apr;7(2):173-9. doi: 10.1007/BF02761753.
As an innovative method for embryo cryopreservation, vitrification not only reduced the cooling stage duration to a minimum, but also eliminated any injuries cased by extracellular ice, which is a major cause of cell injury. Therefore, if embryos are treated adequately, high survival can be obtained. As a component of a vitrification solution, a permeating cryoprotective agent is essential, and additional inclusion of a macromolecule and a small saccharide makes the solution more effective. The author's group composed a solution, designated EFS40, with ethylene glycol, Ficoll, and sucrose. This solution proved effective for the cryopreservation of various stages of embryos in many species. In this article, the author describes the detailed procedure for the vitrification of mouse morulae; related information is also described.
作为一种创新的胚胎冷冻保存方法,玻璃化不仅将冷却阶段的持续时间缩短到最短,还消除了细胞外冰造成的任何损伤,而细胞外冰是细胞损伤的主要原因。因此,如果对胚胎进行充分处理,就能获得较高的存活率。作为玻璃化溶液的一个成分,渗透性冷冻保护剂是必不可少的,额外加入大分子和小分子糖类会使溶液更有效。作者团队用乙二醇、聚蔗糖和蔗糖配制了一种名为EFS40的溶液。该溶液已证明对多种物种不同阶段胚胎的冷冻保存有效。在本文中,作者描述了小鼠桑椹胚玻璃化的详细步骤;同时也介绍了相关信息。