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小鼠MII期卵母细胞的玻璃化冷冻:使用紫杉醇的发育能力

Vitrification of mouse MII oocytes: Developmental competency using paclitaxel.

作者信息

Fesahat Farzaneh, Faramarzi Azita, Khoradmehr Arezoo, Omidi Marjan, Anbari Fatemeh, Khalili Mohammad Ali

机构信息

Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran; Department of Genetics, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.

Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.

出版信息

Taiwan J Obstet Gynecol. 2016 Dec;55(6):796-800. doi: 10.1016/j.tjog.2016.05.010.

Abstract

OBJECTIVE

Oocyte cryopreservation provides an important alternative for fertility preservation for women who will be treated with cytotoxic drugs. However, it can cause spindle disorganization of microtubules, putting the zygote at risk for aneuploidy. Paclitaxel is known to stabilize the microtubules that constitute the spindle. The aim of this study was to investigate the suitable concentration of paclitaxel for adding to the vitrification media to improve the developmental potential of post-thawed mature oocytes to blastocyst formation in mice.

MATERIALS AND METHODS

A total of 300 MII oocytes were retrieved from superovulated mice, and were divided into three groups of control, Experimental I, and Experimental II. Oocytes in Experimental I and Experimental II were cryopreserved in the presence of 0.5μM or 1μM of paclitaxel in vitrification media, respectively. After thawing, all oocytes were incubated in G-IVF medium for 1 hour. From each group,12 oocytes were selected for viability evaluation by Hoechst/propidium iodide nuclear staining. Standard in vitro fertilization was performed on the rest of the oocytes and embryo development was followed to the blastocyst stage.

RESULTS

Fertilization rate was not significantly different between the three groups. However, the cleavage rate (55%) in Experimental II group was significantly lower compared to Experimental I (88%) and control groups (83%). There was a detectable difference between the three groups at the blastocyst rate (Experimental I and control groups, p = 0.004; Experimental II vs. control and Experimental I, p < 0.001). The highest rates of parthenogenesis and arrest were in Experimental II (16% and 21%, respectively) compared with control (6% and 5%, respectively) and Experimental I (5% and 3%, respectively). There was also a significant decrease in viability rate of oocytes in Experimental II compared to the other groups.

CONCLUSION

A high concentration of paclitaxel, an anticancer drug, interrupted the mouse oocyte competency when supplemented to vitrification media. Consequently, the optimal concentration of this cytoskeleton stabilizer may improve the post-thawed developmental abilities of oocytes.

摘要

目的

卵母细胞冷冻保存为接受细胞毒性药物治疗的女性提供了一种重要的生育力保存替代方法。然而,它可导致微管纺锤体紊乱,使合子有非整倍体风险。已知紫杉醇可稳定构成纺锤体的微管。本研究的目的是探究添加到玻璃化液中的紫杉醇适宜浓度,以提高解冻后成熟卵母细胞在小鼠中发育为囊胚的能力。

材料与方法

从超排卵小鼠中获取共300个MII期卵母细胞,并分为对照组、实验组I和实验组II三组。实验组I和实验组II的卵母细胞分别在含有0.5μM或1μM紫杉醇的玻璃化液中冷冻保存。解冻后,所有卵母细胞在G-IVF培养基中孵育1小时。从每组中选取12个卵母细胞,通过Hoechst/碘化丙啶核染色进行活力评估。对其余卵母细胞进行标准体外受精,并跟踪胚胎发育至囊胚阶段。

结果

三组之间的受精率无显著差异。然而,实验组II的卵裂率(55%)显著低于实验组I(88%)和对照组(83%)。三组在囊胚率上存在可检测到的差异(实验组I与对照组,p = 0.004;实验组II与对照组及实验组I相比,p < 0.001)。实验组II的孤雌生殖率和停滞率最高(分别为16%和21%),而对照组分别为6%和5%,实验组I分别为5%和3%。与其他组相比,实验组II的卵母细胞活力率也显著降低。

结论

高浓度的抗癌药物紫杉醇添加到玻璃化液中时会干扰小鼠卵母细胞的能力。因此,这种细胞骨架稳定剂的最佳浓度可能会提高解冻后卵母细胞的发育能力。

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