猕猴精子的冷冻保存以及影响冷冻复苏精子功能的因素
文献类型：学位论文
| 作者 | 司维 |
| 学位类别 | 博士 |
| 答辩日期 | 2004-02 |
| 授予单位 | 中国科学院研究生院 |
| 授予地点 | 北京 |
| 导师 | 季维智 |
| 关键词 | 称猴 精子 冷冻保存 防冻剂 |
| 其他题名 | Cryopreservation of Rhesus Monkey Spermatozoa |
| 中文摘要 | 自从1949年Polge等研究者发现甘油的冷冻保护作用，开创了低温生物学的里程碑以来，精子冷冻保存已经逐渐成为治疗人类不育、家畜繁殖和生物多样性保护的重要手段。然而目前人类对多数物种的基本生殖生物学及其种质冷冻保存的低温生物学知识仍很缺乏。哺乳动物精子冷冻的研究很大程度上是经验性的，即使运用最现代的低温冷冻程序和仪器设备，也仅有约50％的精子能够存活下来。灵长类动物的精子对冷冻非常敏感，迄今为止应用称猴冷冻精子人工受精仅产生过一个后代。为了提高冷冻保存精子的功能和受精能力，本论文以称猴为动物模型，以冷冻复苏精一的运动度、精子质膜和顶体的完整率以及体外受精作为精子功能的检钡l指标，探讨影响动物精子冷冻保存的因素。通过研究1）建立了适合称猴精子冷冻保存的适宜方法以及冷冻复苏精子功能的检测方法，并首次成功地运用冷冻称猴精子体外受精；2）检测了不同甘油浓度和二甲亚飒浓度对冷冻复苏精子运动能力、精子质膜、精子顶体结构和受精能力的影响，确定了称猴精子冷冻保存的适宜甘油浓度；3）对比研究了不同类型渗透性防冻剂在精子冷冻保存中的作用以及对冷冻复苏精子功能的影响，发现对于称猴精子，除了甘油外，乙二醇也是一种优秀的渗透性防冻剂，而丙二醇和二甲亚矾对精子产生了较大的毒害作用，并不适合称猴精子的冷冻保存；4）研究了单糖、二糖和二糖在精子冷冻复苏过程中的保护作用，发现不同类型糖类的冷冻保护作用存在显著差异，单糖和二糖对精子的冷冻保护作用优于二糖，二糖在冷冻复苏过程中对于精子的顶体产生了破坏作用，不同类型的糖类配合使用可以提高对精子的冷冻保护效率。本研究的结果有利于优化灵长类动物精子冷冻保存的方法，包括调整防冻液的成分和冷冻降温程序，从而有助于提高精子冷冻保存的存活率。此外，还有利于深入研究哺乳动物精子冷冻损伤的机制和探讨防冻剂的冷冻保护机理. |
| 英文摘要 | Sperm cryopreservation has provided an effective way for human sterility cure, livestock breeding and biodiversity preservation, since Polge et al (1949) found the cryoprotective effect of glycerol and set up the milestone of cryobiology. However, due to the lack of basic knowledge about reproductive biology and cryobiology of most species, the study of sperm cryopreservation is empirically depended. Even with the most modern freezing protocol and equipments, only about 50% spermatozoa can survive after freezing and thawing. By now, only one offspring of rhesus monkey has been obtained using cryopreserved spermatozoa. The difficulty of cryopreserving nonhuman primate spermatozoa may be due to their fragility. In order to improve the fertility of cryopreserved sperm, factors affecting sperm cryo-survival were studied with rhesus monkey as experimental animal. Sperm motility, plasma membrane integrity, acrosome integrity and in-vitro fertilization were used as functional assessment for frozen-thawed spermatozoa. The present study included: 1) Suitable freezing medium and freezing protocol for rhesus monkey spermatozoa cryopreservation; 2) Effects of different glycerol and dimethyl suifoxide levels on the sperm cryopreservation; 3) Effects of four penetrating cryoprotectants: glycerol, dimethyl sulfoxide, ethylene glycol and propylene glycol on the sperm cryo-survival and function; 4) Cryoprotective roles of monosaccharide, disaccharide and trisaccharide. The Results showed that: 1) rhesus monkey spermatozoa could be successful cryopreservation with TTE; 2) five percent of glycerol is suitable for rhesus monkey sperm freezing; 3) the cryoprotective effects of various penetrating cryoprotectants are different. Dimethyl sulfoxide and propylene glycol are not suitable for rhesus monkey sperm freezing, especially propylene glycol. Ethylene glycol with similar cryoprotective properties to glycerol could be successfully used in the cryopreservation of rheusus monkey spermatozoa; and 4) the cryoprotective effects of various sugar types are different, and combination using of various types of sugar at proper concentration is better than that using one type of sugar alone. The results will be beneficial for spermatozoa cryopreservation of non-human primate and improving the cryo-survival of cryopreserved sperm. |
| 语种 | 中文 |
| 公开日期 | 2010-10-15 |
| 源URL | [http://159.226.149.42:8088/handle/152453/6195]  |
| 专题 | 昆明动物研究所_生殖与发育生物学 |
| 推荐引用方式 GB/T 7714 | 司维. 猕猴精子的冷冻保存以及影响冷冻复苏精子功能的因素[D]. 北京. 中国科学院研究生院. 2004. |
入库方式： OAI收割
来源：昆明动物研究所
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