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近年来,我们初步开展了猪卵透明带(ZP)抗原的提纯工作。在实验过程中,发现ZP 抗原的分离和纯化受许多因素干扰而不够稳定,并且进一步影响到ZP 抗原纯度及活性。为了摸索一个较为适宜的提取条件,我们就ZP 分离纯化的影响因素作一探讨。从屠宰场收集3～5个月刚成熟的猪卵巢,低温状态送到实验室,用自来水多次冲洗并除去结缔组织,进一步用PBS 反复洗涤后置-20℃冻存备用。ZP 制备中采用的过筛及加热等步骤,参照Dun-bar 的报道。猪卵巢的机械剥离用冰冻切片和组织捣碎等方法处理。卵子热处理后,置解剖镜下观察ZP溶解程度,以卵透明带完全溶解,呈现裸卵为标准。各实验组分别计数200～500只卵子,判断透明带完整的卵细胞与裸即的比例,并计算卵透明带溶解的百分率。实验结果如下:一、冰冻切片、外观浓集等对卵子分离的影响:用机械剥离方法破裂卵泡是分离卵子的首要步
In recent years, we initially carried out the zona pellucida (ZP) antigen purification. During the experiment, it was found that the isolation and purification of ZP antigen was disturbed by many factors and not stable enough, and further affected the purity and activity of ZP antigen. In order to explore a more suitable extraction conditions, we discuss the influencing factors of ZP separation and purification. From the slaughterhouse to collect freshly mature pig ovaries 3 to 5 months, sent to the laboratory at low temperatures, washed repeatedly with tap water and remove connective tissue, and then washed repeatedly with PBS and set at -20 ℃ frozen reserve. ZP preparation used in the screening and heating steps, with reference to Dun-bar reported. Mechanical removal of pig ovaries with frozen sections and tissue mashed and other methods. Ovum heat treatment, home dissection microscope to observe the degree of dissolution of ZP to zona pellucida completely dissolved, showing the naked eggs as the standard. The experimental groups were counted 200 ~ 500 eggs, the complete zona pellucida and the ratio of naked that, and calculate the percentage of zona pellucida dissolved. The experimental results are as follows: First, the frozen section, the appearance of concentration on the separation of eggs: the method of mechanical stripping rupture follicles is the first step in the separation of eggs