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目的应用冷冻环、冷冻膜和自制麦管片三种冷冻载体,采用玻璃化冷冻方法冻存昆明小鼠卵裂期胚胎,比较三种载体的冻存效果。方法采集通过体外受精在体外培养第2天得到的4-8细胞期的昆明小鼠胚胎247个,对其中177个按载体不同随机分为冷冻环组59个,冷冻膜组59个,自制麦管片组59个;剩余的新鲜胚胎70个作为对照组,比较冻融后鼠胚的复苏率和囊胚形成率。结果冷冻环组、冷冻膜组和自制麦管片组的胚胎复苏率分别为98.3%、98.3%和94.9%,三组比较差异无统计学意义(P>0.05);冷冻环组、冷冻膜组和自制麦管片组的囊胚形成率分别是84.5%、86.2%和82.1%,三组比较无统计学差异(P>0.05);新鲜对照组囊胚形成率为94.3%,与冷冻环组、冷冻膜组分别比较无统计学差异(P均>0.05),与自制麦管片组有统计学差异(P<0.05)。结论采用玻璃化冷冻方法冻存小鼠卵裂期胚胎,使用冷冻环和冷冻膜作为冷冻载体具有复苏率高、囊胚形成率高的优点,更适合应用于临床。
OBJECTIVE: To freeze the cleavage stage embryos of Kunming mouse by using vitrification method, and to compare the cryopreservation effects of three vectors. Methods Totally 247 Kunming mouse embryos were obtained from in vitro culture on the second day after in vitro fertilization. 177 of them were randomly divided into 5 groups: freezing ring group (59), frozen membrane group (59), self-made wheat 59 slices in the tube group and 70 fresh embryos in the control group. The recovery rate and blastocyst formation rate of frozen embryos were compared. Results The embryo recovery rate of the frozen ring group, the frozen membrane group and the homemade malondialdehyde group were 98.3%, 98.3% and 94.9%, respectively. There was no significant difference between the three groups (P> 0.05) (P> 0.05). The blastocyst formation rate in the fresh control group was 94.3%, which was significantly lower than that in the frozen ring group (P> 0.05). There was a significant difference between the two groups (P <0.05). Conclusion Cryopreservation of the mouse cleavage stage embryos using the freeze-thaw and cryo-membrane as the cryopreservation vector has the advantages of high recovery rate and high blastocyst formation rate, which is more suitable for clinical application.