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The Effect of Nanoparticle on Vitrification of Porcine GV-Stage Oocytes |
1 Institute of Biothermal Technology, University of Shanghai for Science and Technology,Shanghai 200093, China
2 Animal and Veterinary Research Institute,SAAS,Shanghai Municipal Key Laboratory of Agri-Genetics and Breeding,Shanghai 201106,China |
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Abstract Nano-cryopreservation is a promising new way in the next generation of cryopreservation technology; however, using nanoparticles in oocytes vitrification has been rarely reported. This paper investigated the effect of hydroxy apatite (HA), silica, aluminum oxide, and titanium dioxide nanoparticles in the cryoprotectant on the survival rate and developmental rate of porcine GV- stage oocytes. The cells were cryopreserved in Cryotop and observed using fluorescence staining methods. Results showed that HA nanoparticles have the lowest cytotoxicity among the other nanoparticles, the developmental rate of GV-stage porcine oocytes was 100% when concentrate of HA was lower than 0.5%. When the concentration of HA was 0.1% in the cryoprotectant, the developmental rate of GV-stage porcine oocytes was 22% in Cryotop, which was significantly higher than that in the other groups. The size of nanoparticles exerted little influence. When 0.05% HA nanoparticles (60 nm in diameter) were added, the developmental rate increased dramatically from 14.7% in control group to 30.4%. In conclusion, adding appropriate concentration of HA nanoparticles to cryoprotectant can reduce recrystallization during rewarming and promote survival rate and developmental rate of oocytes after freezing and rewarming. The effect of HA nanoparticles is concentration dependent, while independent to diameters of the nanoparticles.
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[1]AlHasani S, Diedrich K, Van der Ven H, et al. Cryopreservation of human oocytes [J]. Human reproduction, 1987, 2(8): 695-700.
[2]郝保同, 刘宝林. 纳米微粒在细胞低温保存中的应用 [J]. 中国组织工程研究与临床康复, 2008, (41): 8140-8142.
[3]徐海峰, 高志新, 刘宝林, 等. 纳米微粒对低温保护剂溶液结晶性质的影响 [J]. 低温与超导, 2010, (11): 53-57.
[4]李方方. 生物材料纳米冷冻过程的理论与实验研究 [D]. 北京:中国科学院研究生院, 2010.
[5]Wang Buxuan, Zhou Leping, Peng Xiaofeng. Viscosity, thermal diffusivity and prandtl number of nanoparticle suspensions [J]. Progress in Natural Science, 2004, (10): 82-86.
[6]Hao Baotong, Liu Baolin. Thermal properties of PVP cryoprotectants with nanoparticles [J]. Journal of Nanotechnology in Engineering and Medicine, 2011, 2(2): 021015.
[7]Kang HU, Kim SH, Oh JM. Estimation of thermal conductivity of nanofluid using experimental effective particle volume [J]. Experimental Heat Transfer, 2006, 19(3): 181-191.
[8]Han X, Ma HB, Wilson C, et al. Effects of nanoparticles on the nucleation and devitrification temperatures of polyol cryoprotectant solutions [J]. Microfluidics and nanofluidics, 2008, 4(4): 357-361.
[9]Wilmut I. The effect of cooling rate, warming rate, cryoprotective agent and stage of development on survival of mouse embryos during freezing and thawing [J]. Life sciences. Pt. 2: Biochemistry, General and Molecular Biology, 1972, 11(22): 1071-1079
[10]韩爽, 李庆宁, 夏天, 等. 医用金属及金属氧化物纳米材料的毒性研究 [J]. 生物物理学报, 2012, 10: 805-814.
[11]吴秋云, 唐萌, 谢彦昕, 等. 不同粒径纳米二氧化硅的体外细胞膜毒性作用 [J]. 中国生物医学工程学报, 2010, 29(3): 437-445.
[12]孟纯阳, 安洪, 蒋电明, 等. 新型纳米骨重建和修复材料羟基磷灰石/聚酰胺体内植入的生物相容性及安全性 [J]. 中国临床康复, 2004, 8(29):6330-6333.
[13]Seki S, Mazur P. Effect of warming rate on the survival of vitrified mouse oocytes and on the recrystallization of intracellular ice [J]. Biology of Reproduction, 2008, 79(4): 727-737. |
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