Comparison and avoidance of toxicity of penetrating cryoprotectants.
Comparison and avoidance of toxicity of penetrating cryoprotectants.
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DOI：
10.1371/journal.pone.0027604
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Eroglu A
中科院分区：
文献类型：
--
作者：
Szurek EA;Eroglu A
本研究的目的是阐明广泛使用的渗透性冷冻保护剂 (CPA) 对哺乳动物卵母细胞的毒性。为此，将小鼠中期 II (M II) 卵母细胞暴露于在含有 10% 胎牛血清的磷酸盐缓冲盐水 (PBS) 中制备的 1.5 M 二甲基亚砜 (DMSO)、乙二醇 (EG) 或丙二醇 (PROH) 溶液。为了解决 CPA 毒性的时间和温度依赖性，将 M II 卵母细胞在室温（RT，~23°C）和 37°C 下暴露于上述 CPA 15 或 30 分钟。随后，通过检查处理后卵母细胞的暴露后存活、受精、胚胎发育、染色体异常和孤雌激活来评估每种 CPA 的毒性。未经处理的卵母细胞作为对照。将 MII 卵母细胞在室温下暴露于 1.5 M DMSO 或 1.5 M EG 15 分钟不会对任何评估标准产生不利影响。相比之下，在相同条件下，1.5 M PROH 诱导卵母细胞变性 (54.2%) 和孤雌激活 (16%) 显着增加。当CPA暴露在37℃下进行时，PROH的毒性作用进一步增加，导致存活率降低（15%）并且不受精，而DMSO和EG的毒性仍然不显着。尽管如此，通过将 PROH 的浓度降低至 0.75 M 并与 0.75 M DMSO 组合，使总 CPA 浓度达到冷冻保护水平，可以完全避免 PROH 的毒性。此外，与单独使用同等浓度的 DMSO 相比，较低浓度（即 0.75 M）的 PROH 和 DMSO 组合显着改善了 MII 卵母细胞的冷冻存活率。综上所述，我们的结果表明，从 CPA 毒性的角度来看，DMSO 和 EG 在缓慢冷却方案中使用更安全，而较低浓度的 PROH 可以与另一种 CPA 组合以避免其毒性并提高冷冻存活率。
The objective of this study was to elucidate the toxicity of widely used penetrating cryoprotective agents (CPAs) to mammalian oocytes. To this end, mouse metaphase II (M II) oocytes were exposed to 1.5 M solutions of dimethylsulfoxide (DMSO), ethylene glycol (EG), or propanediol (PROH) prepared in phosphate buffered saline (PBS) containing 10% fetal bovine serum. To address the time- and temperature-dependence of the CPA toxicity, M II oocytes were exposed to the aforementioned CPAs at room temperature (RT, ∼23°C) and 37°C for 15 or 30 minutes. Subsequently, the toxicity of each CPA was evaluated by examining post-exposure survival, fertilization, embryonic development, chromosomal abnormalities, and parthenogenetic activation of treated oocytes. Untreated oocytes served as controls. Exposure of MII oocytes to 1.5 M DMSO or 1.5 M EG at RT for 15 min did not adversely affect any of the evaluated criteria. In contrast, 1.5 M PROH induced a significant increase in oocyte degeneration (54.2%) and parthenogenetic activation (16%) under same conditions. When the CPA exposure was performed at 37°C, the toxic effect of PROH further increased, resulting in lower survival (15%) and no fertilization while the toxicity of DMSO and EG was still insignificant. Nevertheless, it was possible to completely avoid the toxicity of PROH by decreasing its concentration to 0.75 M and combining it with 0.75 M DMSO to bring the total CPA concentration to a cryoprotective level. Moreover, combining lower concentrations (i.e., 0.75 M) of PROH and DMSO significantly improved the cryosurvival of MII oocytes compared to the equivalent concentration of DMSO alone. Taken together, our results suggest that from the perspective of CPA toxicity, DMSO and EG are safer to use in slow cooling protocols while a lower concentration of PROH can be combined with another CPA to avoid its toxicity and to improve the cryosurvival as well.
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影响因子：
6.1
作者：
GOOK, DA;OSBORN, SM;JOHNSTON, WIH
通讯作者：
JOHNSTON, WIH
DOI：
10.1530/jrf.0.0900547
发表时间：
1990-11-01
期刊：
JOURNAL OF REPRODUCTION AND FERTILITY
影响因子：
--
作者：
CARROLL, J;DEPYPERE, H;MATTHEWS, CD
通讯作者：
MATTHEWS, CD
影响因子：
6.1
作者：
DAMIEN, M;LUCIANO, AA;PELUSO, JJ
通讯作者：
PELUSO, JJ
DOI：
10.1002/mrd.1120160303
发表时间：
1987-03-01
期刊：
GAMETE RESEARCH
影响因子：
--
作者：
GLENISTER, PH;WOOD, MJ;WHITTINGHAM, DG
通讯作者：
WHITTINGHAM, DG
影响因子：
2.7
作者：
Arav, A;Zeron, Y;Crowe, JH
通讯作者：
Crowe, JH