Universidade do Vale do Itajaí (UNIVALI), Laboratory of Reproductive Biology, Itajaí/SC, Brasil.
Fertas Brasil, Balneário Camboriú/SC, Brasil.
JBRA Assist Reprod. 2021 Apr 27;25(2):223-228. doi: 10.5935/1518-0557.20200084.
To evaluate in vitro oocyte maturation rates in embryonic culture medium after induction by hyperosmotic shock caused by exposure to vitrification solutions.
Bilateral oophorectomy was performed on 20 prepubescent female mice (Swiss). Immature (Prophase I) oocytes (N = 400) were obtained by ovarian dissection, divided into 4 groups, and transferred to culture dishes containing fertilization medium (Sydney IVF Fertilization Medium, Cook® Medical). The control group (CG) did not receive treatment, the test groups (G1, G2, G3) were treated with vitrification solution - 2 (VI-2: 14 M sucrose + ethylene glycol and dimethyl sulfoxide) for 30 seconds and subsequently: G1: 30 seconds in devitrification solution - 2 (DV-2: 0.5M sucrose); G2: 60 seconds DV-2; G3: 60 seconds DV-1(1M sucrose) and 180 seconds DV-2. All groups were cultivated for 24 hours in an incubator at 37ºC and 5% CO2 (Thermo model 3110). After this period, we checked their maturation status.
Oocytes exposed to VI-2, DV-1 and DV-2 (G3) showed the highest rate of competence in resuming meiosis and reaching the MII stage; however, there was no statistically significant difference (G3 = 50.5% - 49/97; CG = 27.8% - 10/30).
Oocyte exposure to vitrification solutions, in order to cause osmotic shock, did not interfere with the resumption of meiosis in mice oocytes.
评估玻璃化溶液诱导的高渗休克对胚胎培养液中卵母细胞体外成熟率的影响。
对 20 只未成熟(前期 I 期)的雌性小鼠(瑞士)进行双侧卵巢切除术。通过卵巢解剖获得不成熟(前期 I 期)卵母细胞(N=400),将其分为 4 组,转移到含有受精培养基(悉尼 IVF 受精培养基,库克®医疗)的培养皿中。对照组(CG)未进行处理,实验组(G1、G2、G3)用玻璃化溶液-2(VI-2:14M 蔗糖+乙二醇和二甲基亚砜)处理 30 秒,随后:G1:在脱玻璃化溶液-2(DV-2:0.5M 蔗糖)中处理 30 秒;G2:在 DV-2 中处理 60 秒;G3:在 DV-1(1M 蔗糖)中处理 60 秒和 DV-2 中处理 180 秒。所有组均在 37°C 和 5%CO2(Thermo 型号 3110)的孵育箱中培养 24 小时。在此期间,我们检查了它们的成熟状态。
暴露于 VI-2、DV-1 和 DV-2(G3)的卵母细胞恢复减数分裂并达到 MII 期的能力最高;然而,这并没有统计学上的显著差异(G3=50.5%-49/97;CG=27.8%-10/30)。
为了引起渗透休克,玻璃化溶液暴露并没有干扰小鼠卵母细胞减数分裂的恢复。