The influence of cryopreservation on murine oocyte water permeability and osmotically inactive volume
The influence of cryopreservation on murine oocyte water permeability and osmotically inactive volume
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DOI：
10.1006/cryo.1996.1984
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发表时间：
1997-02-01
期刊：
影响因子：
2.7
通讯作者：
Fuller, B
中科院分区：
文献类型：
--
作者：
Litkouhi, B;Marlow, D;Fuller, B
对未冷冻(N=18)和冷冻(N=21)的ICR小鼠卵母细胞进行了渗透实验，以确定标准的冷冻程序是否改变了膜透性(渗透系数，L(P))和/或渗透不活化量(V-b)。卵母细胞最初在等渗(288mOsm)的氯化钠溶液中，在微扩散室内暴露于900mOsm的氯化钠中。细胞大小的变化被录像，并使用参数估计程序进行分析。本文首次给出了以渗透不活度体积作为拟合参数的双参数模型(L(P)和V-b)的最佳估计。未冷冻对照种群的平均L(P)和平均V-b值(L(P)=0.64+/-0.15mum/min/atm，V-b=24.7+/-2.9%)与冷冻保存种群(L(P)=0.63+/-0.12mum/min/atm，V-b=28.0+/-10.8%)之间无显著差异。而冷冻保存过程对L(P)、V-b的平均值和L(P)的变异性没有显著影响，但却产生了更多的变异性。超低温保存产生的V-b变异性增加的原因尚不清楚。这些结果表明，未冷冻对照卵母细胞的渗透特性可以作为冷冻解冻卵母细胞的一个合理的近似值。他们还表明，多参数模型和参数估计方法可能有助于更全面地了解在这里检测到的渗透特性的更细微的变化。统计检验也首次被用来证实所有实验总体都来自正态分布的假设。(C)1997年学术出版社。
Osmotic experiments were performed on unfrozen (N = 18) and cryopreserved (N = 21) ICR murine oocytes in order to determine whether a standard cryopreservation process alters membrane water permeability (hydraulic conductivity, L(p)) and/or osmotically inactive volume (V-b). Oocytes, initially in an isotonic (288 mOsm) NaCl solution, were exposed to 900 mOsm NaCl in a microdiffusion chamber. Cell size changes were videotaped and analyzed using a parameter estimation program. Best estimates for a two-parameter model (L(p) and V-b) which includes the osmotically inactive volume as a fitting parameter are presented for the first time. The cryopreservation process produced no significant difference between the mean L(p) or the mean V-b values for the unfrozen control population (L(p) = 0.64 +/- 0.15 mu m/min/atm, V-b = 24.7 +/- 2.9%) and the cryopreserved population (L(p) = 0.63 +/- 0.12 mu m/min/atm, V-b = 28.0 +/- 10.8%). While the cryopreservation process did not cause significant changes in the mean values of L(p), V-b, or the variability of L(p), it did produce more variability of V-b. The cause of the increased variability of V-b produced by cryopreservation is unknown. These results suggest that the osmotic properties of unfrozen control oocytes can be used as a reasonable approximation for frozen-thawed oocytes. They also suggest that multiple parameter models and parameter estimation methods may be useful in developing a more comprehensive understanding of the more subtle alterations in osmotic properties that were detected here. Statistical tests were also used for the first time to confirm the assumption that all of the experimental populations were derived from normal distributions. (C) 1997 Academic Press.