超速冷冻法冷冻保存大鼠卵巢组织的实验研究

Study on the morphology and function after ultrarapid cryopreservation of ovarian tissue in rats

目的探讨超速冷冻法对SD大鼠卵巢组织的冷冻保存功效。方法采用液氮直投法冷冻大鼠卵巢组织。计数冻融后卵巢组织内形态正常始基卵泡数量。将卵巢组织自体移植,术后35天处死动物,测定血清雌二醇(E2)水平。结果新鲜和冻融后的大鼠卵巢组织中形态正常始基卵泡数量无统计学差异(P>0.05)。移植术后35天,两移植组血清E2水平无统计学差异(P>0.05)。结论液氮直投超速冷冻法可以有效的保存卵巢组织,对大鼠卵巢组织学和功能的影响较小。

Objective： To observe the effects of cryopreservation on morphology and function of rats ovarian tissues after ultrarapid freezing.Methods： Cryopreservation of rats ovary pieces was accomplished by direct plunging into liquid nitrogen.Histological sections of fresh and cooled ovarian pieces were analyzed and the number of viable primordial follicles was counted.Fresh and frozen-thawed ovarian pieces were autotransplanted heterotopically（in the back subcutis） to the bilaterally ovariectomized adult female SD rats. After transplantation,the serum levels of E2 were determined.Results： The number of normal primordial follicles in fresh and cryopreserved tissue samples was not significantly different.The results of serum estradiol showed no significant difference（P〉0.05） after transplantation in 35 days between the frozen-thawed group and the fresh group or the sham group.Conclusion： Cryopreservation by direct plunging pieces into liquid nitrogen is an effective way to preserve rats＇ ovarian pieces.