Spindle organization after cryopreservation of mouse, human, and bovine oocytes

Oocyte cryopreservation would alleviate a number of ethical, social, and religious problems associated with human embryo storage. One potential problem is the effect of cryopreservation on the metaphase II spindle and chromosomes. The microtubules that make up the spindle tend to depolymerize at sub-physiological temperatures. Although there are numerous reports in the literature on this topic, discrepancies as to whether the spindle can or cannot reform persist. One of the confounding factors may be the low cryosurvival rates (around 50%) for mammalian oocytes. In recent years, a cryopreservation medium and protocol have been developed that allow oocytes of several species to be cryopreserved with high survival rates (>85%). Bovine, mouse, and human oocytes consistently reformed a morphologically normal spindle with chromosomes aligned along the metaphase plate (70% or higher) after first surviving cryopreservation (>87% survival for all species tested). Normal chromosome numbers were found in every second polar body tested by FISH (second polar bodies n = 4). It is concluded that the mammalian spindle, although depolymerized during cryopreservation, has the ability to reform, and in the mouse has been shown to function normally. Therefore, spindle reformation may not be a major cause for concern when storing mammalian MII oocytes.