目录
摘要II
关键词II
AbstractIII
引言
引言1
1材料与方法1
1.1猪卵巢的处理1
1.2主要试剂及器材 1
1.3试验方法 1
1.3.1观察小卵1
1.3.2采集和初次分离小卵2
1.3.3进一步分离纯化方法2
1.3.4记录采集数量2
2结果与分析2
2.1小卵的观察2
2.2初次分离的到的小卵3
2.3不同方法进一步分离小卵液得到的小卵4
2.3.1胰酶离心消化法 4
2.3.2口吸管法4
2.3.3细胞过滤筛法5
2.3.4采集数量的记录6
3讨论 7
致谢10
参考文献12
对猪卵巢内未形成透明带前卵母细胞分离方法的研究
摘 要
[目的]本试验旨在对从屠宰场运回的猪卵巢内未形成透明带前卵母细胞（小卵）的分离进行可行性验证。[方法]从带有卵母细胞核卵丘细胞的卵泡中抽取卵泡液，随后在显微镜下用口吸管分离、细胞分离筛法、离心胰酶消化法这三种方法分别进一步分离未形成透明带前卵母细胞。[结果]结果表明口吸管法效果最好，分离得到的小卵纯度高，但分离效果受个人操作经验影响较大。胰酶离心法效果相对较差，分离出的卵母细胞数量较少，而且混有大量的颗粒细胞，我们认为需要进一步实验对参数和分离液进行对比调试。此外，虽然颗粒细胞直径小于40μm，但由于颗粒细胞团的存在，小卵液中仍存在着大量的颗粒细胞，因此细胞分离筛不能有效地对小卵进行分离。[结论]口吸管法可有效分离小卵（未形成透明带前卵母细胞），另外两种方法对小卵的分离还需要进一步的研究。
THE METHODS RESEARCH ON OF THE ISOLATION OF PIG OOCYTES BEFORE ZONA PELLUCIDA FORMATION
ABSTRACT
[Objectives] The purpose of this study is to verif *51今日免费论文网|www.51jrft.com +Q: *351916072* 
y the feasibility of the separation of oocytes before the zona pellucida (ZP) formation (small eggs) from pig ovaries which collected from slaughter house. [Methods] Follicular fluid were aspirated from the follicle with oocytes and cumulus cells. After that, oocytes which have not been formed ZP were sorted by three methods, mouth pipette selection under the microscope, cell filter and digestioncentrifugation. [Results] Results showed that the method of mouth pipette selection was the best method, and the purity of small eggs was high. However, the separation effect is greatly affected by personal because for the personal handling experience. The effect of digestioncentrifugal is relative lower, the number of sorted oocytes were less but mixed with a large number of granular cells. It is speculated that the parameters for centrifugation are need more test. In addition, although the diameter of granular cells is less than 40 μm, the granular cells are still remained with the small eggs because of granular cells. Therefore, the small eggs can not be efficient sorted by the cell filter. [Conclusions] In conclusion, small eggs (oocytes which has not formed the ZP yet) can be effectively separated by using the mouth pipette. More studies are needed for sorting the small eggs by the other two methods.
KEY WORDS: oocyte; pig; zona pellucida; mouth pipette; cell fiter

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