母核

It is concluded that (1) there are no relationships between fusion rate and relativeness of the recipient cytoplasm to nucleus donor cells,(2) cytoplast of the goat MII oocyte can support the preimplantation development of SCNT embryos reconstructed with nucleus from other species,(3) the blastocyst rate of close relative inter-species SCNT embryos is higher than that of distant relative inter-species SCNT embryos.

由此结果得出以下结论：(1)山羊M II期卵母细胞胞质与供核细胞之间的亲缘性不影响两者的融合率；(2)山羊M II期卵母细胞的胞质能支持异种间体细胞核移植胚的着床前发育；(3)亲缘关系近的种间核移植胚的囊胚发育率高于亲缘关系远的种间核移植胚的。

The porine is the polyembryony animal, many obtains the ovocyte from the porine ovary although but the maturity quite to be low; Because the ROSI technology is does not have to grow the mature sole round sperm cell to pour into directly completely in the ovicell nature, jumped over the spermatozoon in to pass through the physiology and the biochemistry, if the ovicell nature mature or the activation degree were insufficient, added the round immature sperm cell maturity quite inferior reason, very possibly Causes the ROSI micro fertilization defeat.

猪是多胎动物，从猪卵巢上获得的卵母细胞数量虽然较多，但成熟度比较低；由于ROSI技术是将没有完全发育成熟的单一圆形精细胞直接注入卵胞质内，跳越了精子在穿过透明带和卵质膜等过程中所发生的生理和生化反应；如果卵胞质成熟或活化程度不够，加之圆形未成熟精细胞成熟度比较差等原因，很可能造成ROSI受精的卵母细胞内部激活因子蓄积减少和生成不足，引起精核解聚困难、精圆核不能形成、第二极体不能排出、卵母细胞孤雌发育率增加等，使ROSI显微受精失败。

The cleavation rates and blastocyst development rates of cloned embryos were used to assess the efficiency of different operational procedure. Finally, the best combination of operational procedure, that the spindle-viewer system was used for oocytes enucleating, and donor cell was electrofused into ooplasm by electrical pulse (1.9 kV/cm, 10 ms, two) to reconstruct bovine cloned embryos.

以核移植胚胎的卵裂率、囊胚发育率作为检测指标，对不同的方法所获得的克隆胚胎的卵分裂率与囊胚发育率进行比较，最后筛选获得1个优化的牛体细胞核移植操作程序，即采用Spindle view系统对牛卵母细胞进行去核操作，将供核体细胞注射到卵周隙，然后通过电融合法将供体核引入去核卵细胞质(电融合参数为1.9 kV/cm，脉冲时程10 ms，方波2次间隔2 s)。

In conclusion, these results suggested that (1) the ageing-associated decline in fertility of female KM mouse was due to a decrease in both quantity and quality of oocytes;(2) the ooplasm from young mice did not correct the meiotic errors of aged mice, and the ooplasm from aged mice did not induce abnormal segregation of meiotic chromosome of young mice, indicating that meiotic anomalies found in the oocytes of aged mice might be related to nucleus or chromosomes and relevant factors;(3) a critical nucleocytoplasmic ratio was essential for normal maturation and segregation of meiotic chromosomes of oocytes and development to 2-cell embryos.

本研究结果表明：（1）昆明白小鼠与衰老相关的生育力下降是其卵母细胞数量减少和质量下降的综合结果；（2）年轻小鼠卵母细胞细胞质不能纠正老龄小鼠GV的减数分裂错误，老龄小鼠卵母细胞细胞质也不诱导年轻小鼠的GV发生减数分裂错误，老龄小鼠卵母细胞减数分裂异常很可能与细胞核或染色体及其相关因素有关；（3）关键的核质比对卵母细胞正常的成熟、减数分裂染色体分离及2-细胞期胚的发育是绝对必需的。

Methods A C57BL/6j mouse cumulus cell nucleus 10-12 mm in diameter was inserted into the perivitelline space of an enucleated oocyte.

方法将直径10~12mm的C57BL/6j小鼠卵丘细胞核注射到去核卵母细胞透明带下，构建供体核-卵母细胞复合体。

The situation of abnormal development of male cells is as follows:microspore mother cell can't enter into meiosis because of intense vacuolation,shrink and disintegration of its cytoplasm;although vacuolated microspore mother cell can enter into meiosis,it can't form normal dyad and degenerate in the middle process;dyad and tetrad become vacuolated and can't develop normally;cytoplasm of microspore shrinks around the nucleus at the stage of central nucleus microspore,the shape of microspore is twisted into crescent or irregular shape,at last its cytoplasm and nucleus are disintegrated and crescent vacant microspore presents;nutritive substances can't be accumulated at the stage of vacuolated microspore,cytoplasm is disintegrated,and microspore turns into a big vacant pollen.

雄性细胞异常发育有几种情况：小孢子母细胞强烈液泡化，细胞质收缩解体，不能进入减数分裂；小孢子母细胞液泡化，虽能进入减数分裂，但不能形成正常二分体而中途退化；二分体、四分体细胞液泡化，不能进行正常发育；单核小孢子中央期，细胞质收缩包围核，小孢子形状扭曲呈月牙形或不规则形，最终细胞质和核解体而呈月牙形的空壳小孢子；单核液泡期的小孢子不能积聚营养物质，细胞质解体而成为大的空壳花粉粒。

In order to establish mouse nuclei transfer technology, mouse zygote culture medium and activated conditions were optimized in vitro and enucleated method of MII oocyte were investigated in this study.

本研究通过对小鼠受精卵体外培养体系，MII期卵母细胞孤雌激活条件的筛选以及探讨MII期卵母细胞的去核方法为小鼠体细胞核移植的成功奠定基础。

In order to find an effective nuclear transfer program, this thesis investigated three factors affecting the efficiency of somatic nuclear transfer on goats: the in vivo mature time of oocytes, the component of operation liquid for oocyctes enucleating and the parthenoactivation mode of oocytes.

本实验通过改变以下试验条件：卵母细胞体外成熟时间的不同、卵母细胞去核时操作液成分的不同，来研究其对山羊核移植效率的影响，希冀寻找一个有效的核移植方案。

A new method for enucleating MⅡ nucleus in mouse and rat was induced; Living births were obtained from IVF of mouse oocytes reconstructed by transfer of metaphase Ⅱ chromosomes. For the interspecies nuclear transfer between mouse and rabbit, the reconstructed embryos could develop into blasocyst in vitro.

在不同品系小鼠卵母细胞核质交换移植中将电融合的重构卵进行IVF，经胚胎移植后顺利产仔；在异种核移植方面，小鼠核与兔卵母细胞的异种重构胚在体外培养条件下发育至囊胚阶段。

In the present study, we collected cumulus cells oocyte complex from ovaries of two different strain mice. The cumulusenclosed oocytes were cultured for 6 h in MEM supplemented with growth factor and FSH. The meiotic maturation of these oocytes has progressed to pro-metaphse Ⅰ stage and the condensed chromosomes are visible under DIC microscope, metaphase Ⅰ spindle even can be detected under Polscope. The metaphase Ⅰ spindles of oocytes were exchanged under such microscopes. After electric stimuli, 91. 6% and 91. 6% karyoplasts-cytoplasm pairs were fused respectively. The resulting oocytes were cultured further in MEM and over 80% of oocytes released the first polar body. 79% and 77% of oocytes formed two pronuclei after in vitro fertilization and the embryos were cultured in KSOM supplemented with amino acids. Over 60% of embryos developed to blastocyst stage.

在本研究中我们在取得两种不同品系小鼠的卵丘卵母细胞复合体后，先将卵丘卵母细胞复合体置于含有多种生长因子和激素的MEM培养液中培养6小时，此时卵母细胞已进入第一次减数分裂的前中期，并且在DIC倒置显微镜下可以看到浓缩的染色体，用Polscope可以发现明显的纺锤体，借助这种显微镜通过显微操作将两种不同品系小鼠来源的卵母细胞的MI纺锤体进行互换，经过三次直流电脉冲作用后，分别有91.6％的胞质—MI核质体对融合，经过进一步的培养后，超过80％的重组卵母细胞排出第一极体，体外受精后分别有79％和77％的重组卵形成双原核，受精后的胚胎在KSOM胚胎培养液中体外培养4天后，超过60％的胚胎发育至囊胚。

This one mode pays close attention to network credence foundation of the businessman very much.

这一模式非常关注商人的网络信用基础。

Cell morphology of bacterial ghost of Pasteurella multocida was observed by scanning electron microscopy and inactivation ratio was estimated by CFU analysi.

扫描电镜观察多杀性巴氏杆菌细菌幽灵和菌落形成单位评价遗传灭活率。