不同浓度保护剂对超速冻存小鼠卵巢组织的效果观察

目的:寻找适宜的小鼠卵巢组织超速冻存低温保护剂种类及浓度。方法:以不同浓度二甲亚砜(DMSO)和丙二醇(PROH)(1.5　mol/L、3　mol/L)作低温保护液时,观察超速冻存法对小鼠卵巢组织的冻存效果。解冻后行自体和异体卵巢的肾被膜下移植。通过对受体动情期的恢复及激素水平测定等方法对冻存效果进行观察。结果:低浓度组DMSO和PROH动情期的恢复率分别为75%和71%;血清E2分别为(10.5和16.8)×10-9　mmol/L,与作为对照的慢速程序法组无显著性差异(66.7%和100%;11.7×10-9　mmol/L和12.1×10-9　mmol/L)。高浓度动情期的恢复率分别为100%和62.5%;血清E2分别为14.1×10-9　mmol/L(DMSO)和7.4×10-9　mmol/L(PROH),与慢速组比也无显著性差异。结论:两种浓度的DMSO和PROH均适用于超速冻存法,尽管高浓度的DMSO在超速冻存时可取得较好的动情周期恢复率,但血清E2 水平的结果以低浓度的PROH为好。

Effects of Cryopreservation of Different Concentration Cryoprotective Agents in Ultrarapid Freezing Mice Ovarian Tissue

Objective:To observe the effects of two types of cryoprotective agents (CPA) of different concentration on mice ovarian tissue by ultrarapid freezing and thawing. Methods: Mice ovarian tissues were cryopreserved in a low concentration freezing solution consisting of 1.5 mol/L DMSO or propanediol (PROH) and 0.1 mol/L sucrose and a high concentration freezing that consisting of 3.0 mol/L DMSO or PROH and 0.5 mol/L sucrose by ultrarapid freezing method(UFM). At the same time, the slow procedure freezing method(SPFM) was used as control method using low concentration freezing solution. The effect of cryoprotection were estimated by observing the recovery rate of estrus, levels of serum estrogen of recipient mice after tissue transplantation under kidney capsule. Results: In the low concentration group, the recovery rate of estrus cycle was 75% and 71% respectively, the levels of serum E2 was 10.5×10-9mmol/L and 16.8×10-9mmol/L respectively, similar to that of SPFM (66.7% and 100%; 11.7×10-9mmol/L and 12.1×10-9 mmol/L). In the high concentration group, the recovery rates of estrus cycle were 100% and 62.5% respectively; the level of serum E2 was 14.1 and 7.4 × 10-9 mmol/L, respectively, there were no significant differences compared with that of SPFM, either. Conclusion: These results suggest that by ultrarapid freezing method either two concentrations of CPA could effectively preserve mice ovarian tissue; although the recovery rate of estrus cycle was very good with 3.0 mol/L DMSO, the higher level of serum E2 can be obtained with 1.5 mol/L PROH.