羟基磷灰石纳米颗粒提高了玻璃化猪卵母细胞的存活率及其机制。

Hydroxyapatite nanoparticles improved survival rate of vitrified porcine oocytes and its mechanism.

作者信息

Zhou Xinli, Li Weijie, Zhang Defu, Dai Jianjun

出版信息

Cryo Letters. 2015 Jan-Feb;36(1):45-50.

Abstract

BACKGROUND

Cryopreservation of oocyte has become most essential for the long-term conservation and widespread dispersion of animal genetic resources, but the efficiency of oocyte cryopreservation remains low.

OBJECTIVE

The present study is to study the effect of hydroxyapatite (HA) nanoparticles on the survival rate of vitrified oocytes and the possible mechanism of cryopreserving oocytes with nanoparticles.

MATERIALS AND METHODS

Porcine oocytes were vitrified in cryoprotectant (CPA) with biocompatible HA nanoparticles by Cryotop. The recrystallization of nano-CPA was observed with cryomicroscope.

RESULTS

When 0.01%, 0.02%, 0.05% and 0.1% HA nanoparticles were added into vitrification solution, the survival rate of oocytes after in vitro maturation ranged from 25.9% to 35.4%, which is significantly higher than group without HA nanoparticles (14.7%). The microphotographs of oocytes in different solutions during freezing, thawing and melting showed that HA nanoparticles at a certain concentration can hinder the recrystallization of vitrification solution during rewarming.

CONCLUSION

We speculate that preventing the recrystallization is the reason that nano-CPA can promote the survival rate of vitrified oocytes.

摘要

背景

卵母细胞的冷冻保存对于动物遗传资源的长期保存和广泛传播已变得至关重要，但卵母细胞冷冻保存的效率仍然很低。

目的

本研究旨在探讨羟基磷灰石（HA）纳米颗粒对玻璃化卵母细胞存活率的影响以及纳米颗粒冷冻保存卵母细胞的可能机制。

材料与方法

采用Cryotop法，将猪卵母细胞在含有生物相容性HA纳米颗粒的冷冻保护剂（CPA）中进行玻璃化。用低温显微镜观察纳米CPA的重结晶情况。

结果

当向玻璃化溶液中添加0.01%、0.02%、0.05%和0.1%的HA纳米颗粒时，体外成熟后卵母细胞的存活率在25.9%至35.4%之间，显著高于未添加HA纳米颗粒的组（14.7%）。不同溶液中卵母细胞在冷冻、解冻和融化过程中的显微照片显示，一定浓度的HA纳米颗粒可在复温过程中阻碍玻璃化溶液的重结晶。