The effect of osmotic stress on the metaphase II spindle of human oocytes, and the relevance to cryopreservation.
The effect of osmotic stress on the metaphase II spindle of human oocytes, and the relevance to cryopreservation.
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DOI：
10.1093/humrep/deh201
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发表时间：
2004-05-01
影响因子：
6.1
通讯作者：
Critser, JK
中科院分区：
文献类型：
--
作者：
Mullen, SF;Agca, Y;Critser, JK
背景：了解渗透压耐受限度对于设计最佳的细胞冷冻保存程序非常重要。方法：将成熟的人卵母细胞在37 ℃下暴露于浓度为35、75、150、600、1200或2400（+/-5）毫渗摩尔（mOsm）的各向异性蔗糖溶液中。还使用290 mOsm的对照处理。将卵母细胞随机分配至各实验处理组。处理后，将卵母细胞培养1 h，然后在冷甲醇中固定。免疫细胞化学染色和荧光显微镜用于评估中期II（MII）纺锤体的形态。逻辑回归用于确定培养基渗透压摩尔浓度是否对纺锤体结构具有显著影响。结果：渗透压是纺锤体形态的重要预测因子。在35，75，和150 mOsm的低渗效应导致100，67，和56%的卵母细胞异常纺锤体，分别。在600，1200和2400 mOsm的高渗作用导致44，44和100%的纺锤体结构异常，分别。结论：等渗条件导致人卵母细胞MII纺锤体破坏。将这一基本知识应用于人类卵母细胞冷冻保存应导致维持活力的细胞数量增加。
BACKGROUND: Knowing osmotic tolerance limits is important in the design of optimal cryopreservation procedures for cells. METHODS: Mature human oocytes were exposed to anisosmotic sucrose solutions at concentrations of 35, 75, 150, 600, 1200, or 2400 (+/-5) milliosmolal (mOsm) at 37degreesC. A control treatment at 290 mOsm was also utilized. Oocytes were randomly allocated to each experimental treatment. After the treatment, the oocytes were cultured for 1 h, then fixed in cold methanol. Immunocytochemical staining and fluorescence microscopy were used to assess the morphology of the metaphase II (MII) spindle. Logistic regression was used to determine if media osmolality had a significant effect on spindle structure. RESULTS: Osmolality was a significant predictor of spindle morphology. Hyposmotic effects at 35, 75, and 150 mOsm resulted in 100, 67, and 56% of oocytes having abnormal spindles, respectively. Hyperosmotic effects at 600, 1200, and 2400 mOsm resulted in 44, 44, and 100% of the spindles with abnormal structure, respectively. CONCLUSIONS: Anisosmotic conditions lead to disruption of the MII spindle in human oocytes. Applying this fundamental knowledge to human oocyte cryopreservation should result in increased numbers of cells maintaining viability.