The effect of osmotic stress on the cell volume, metaphase II spindle and developmental potential of in vitro matured porcine oocytes
The effect of osmotic stress on the cell volume, metaphase II spindle and developmental potential of in vitro matured porcine oocytes
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DOI：
10.1016/j.cryobiol.2007.03.005
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发表时间：
2007-06-01
期刊：
影响因子：
2.7
通讯作者：
Critser, J. K.
中科院分区：
文献类型：
--
作者：
Mullen, S. F.;Rosenbaum, M.;Critser, J. K.
猪的动物模型被用来推进我们对人类生理学的理解。目前的研究还针对生产转基因猪的方法。冷冻库配子和胚胎可以促进有价值的基因型的保存，但冷冻保存猪卵母细胞已被证明是非常具有挑战性的。本研究旨在了解不等渗溶液对体外成熟猪卵母细胞的影响，作为设计改进冷冻保存程序的第一步。我们假设，卵母细胞的比例，表现出正常的纺锤体装置和体外发育潜力将成比例的溶液渗透压。将卵细胞在38 ℃下在各种低渗或高渗溶液和等渗对照溶液中孵育10分钟，然后评估这两个参数。我们的研究结果支持这一假设，随着不等张暴露水平与等张暴露水平的偏离，纺锤体的比例越来越大。与未处理的对照组相比，暴露于不等渗溶液后，只有约一半的卵母细胞保持发育潜力。卵母细胞体积显示出线性响应的非等渗溶液的预期，估计相对无活性细胞体积为0.178。本研究的结果为猪卵母细胞的生物物理特性提供了初步的数据。从未来的实验设计，以确定膜渗透性的各种冷冻保护剂的结果将允许预测建模的最佳冷冻保存参数，并提供了一个基础，设计改进的冷冻保存程序。(c)2007爱思唯尔公司All rights reserved.
Porcine animal models are used to advance our understanding of human physiology. Current research is also directed at methods to produce transgenic pigs. Cryobanking gametes and embryos can facilitate the preservation of valuable genotypes, yet cryopreserving oocytes from pigs has proven very challenging. The current study was designed to understand the effects of anisotonic solutions on in vitro matured porcine oocytes as a first step toward designing improved cryopreservation procedures. We hypothesized that the proportion of oocytes demonstrating a normal spindle apparatus and in vitro developmental potential would be proportional to the solution osmolality. Cocytes were incubated for 10 min at 38 degrees C in various hypo- or hypertonic solutions, and an isotonic control solution and then assessed for these two parameters. Our results support the hypothesis, with an increasing proportion of spindles showing a disrupted structure as the levels of anisotonic exposure diverge from isotonic. Only about half of the oocytes maintained developmental potential after exposure to anisotonic solutions compared to untreated controls. Oocyte volume displayed a linear response to anisotonic solutions as expected, with an estimated relative osmotically inactive cell volume of 0.178. The results from this study provide initial biophysical data to characterize porcine oocytes. The results from future experiments designed to determine the membrane permeability to various cryoprotectants will allow predictive modeling of optimal cryopreservation parameters and provide a basis for designing improved cryopreservation procedures. (c) 2007 Elsevier Inc. All rights reserved.