卵裂细胞

It showed that the cumulus cells did play significant role in the stimulation of nuclear maturation of porcine oocytes and subsequent development.

表明卵丘细胞对猪卵母细胞的核成熟及卵母细胞体外受精后的卵裂和早期胚胎发育，都具有明显的促进作用。

These observations show that reprogramming is easier in interspecific embryos reconstructed with ES cells than that reconstructed with somatic cells, and that ES cells have the higher ability to direct the reconstructed embryo development normally than fibroblast cells. Oocytes were reconstructed with outbreeding Kunming albino mouse ES cells and enucleated rabbit oocytes, and the effects of the passages of ES cells and 6-DMAP on the development of interspecific reconstructed oocytes were analyzed. The interspecific reconstructed ES-rabbit oocytes were activated either by combined two set electric pulses and 6-DMAP or by two set electr

以上结果显示，6－DMAP能增加胚胎干细胞异种重构卵的卵裂率，对重构卵囊胚的发育率影响不大；高培养代数和低培养代数的胚胎干细胞用于异种核移植不影响异种重构卵的卵裂率和囊胚发育率；用胚胎干细胞作为供核细胞；比体细胞作为供核细胞所构建的异种重构胚更容易进行重编程，并且胚胎于细胞指导异种克隆胚胎正常发育的能力强于体细胞。

Different strains have different reaction to the delay of oviducts in body after animal death. After 10min delay C57BL/6 mouse oocytes have death rate of 56.5%,significantely higher than Kunming mouse (47.6%);Spontaneous activation rates were respectively 13.3% and 46.0%, C57BL/6 were obviously lower than Kunming mouse.4. The oviducts were obtained after being delayed 5min 24h after the mice were injected with hCG. The oocytes were cultured in CZB. About 81.1% occurred spontaneous activation, evidently lower than parthenogenetic rate (96.4%) with SrCl_2. Spontaneous activable oocytes had high cleavage rate(93.2%) and 4-cell rate(87.3%). However, spontaneous activable oocytes had blastula development rate(18.7%) as low as parthenogenetic oocytes by SrCl_2(22.9%).

不同品系小鼠卵母细胞对输卵管在体内滞留产生的反应不同，滞留10min C57BL/6系小鼠卵母细胞死亡率为56.5%，显著高于昆明鼠(47.6%)；自发激活率分别为13.3%和46.0%，C57BL/6系小鼠显著低于昆明鼠。4.hCG后24h体内滞留5min卵母细胞在CZB中培养自发激活率为81.1%，显著低于SrCl_2孤雌激活率(96.4%)；自发激活的卵母细胞有较高的卵裂率(93.2%)和4-cell比率(87.3%)，但囊胚率(18.7%)较低，同卵龄的卵母细胞经SrCl_2孤雌激活囊胚发育率为22.9%，差异不显著。

When electric fusion method was used for nuclear transfer, the fusion rate (46. 0%), cleavage rate (53. 9%) and blastocyte development rate (10.9%) of adult ear fibroblasts were significantly lower than that of fetal fibroblasts (64. 5%, 70.1%, 21. 6% respectively), fetal skin cells (71. 5%, 70.8%, 22. 1% respectively) and ovary granulosa cells (88. 2%, 79. 1%, 25. 5% respectively). There was no significant difference among other donor cells in the cleavage and blastocyst development rate of resconstituted embryos.

当用电融合法进行核移植时，成体耳部成纤维细胞的融合率（46.0％），卵裂率（53.9％）和囊胚发育率（10.9％）均显著低于胎儿成纤维细胞（64.5％，70.1％和21.6％），胎儿皮肤细胞（71.5％，70.8％和22.1％），以及卵巢颗粒细胞（88.2％，79.1％和25.5％）；另外三种细胞间的卵裂率，囊胚发育率无显著差异，但卵巢颗粒细胞的融合率显著高于胎儿成纤维细胞和胎儿皮肤细胞（88.2％vs 64.4％，71.5％，P＜0.05）。

The results showed that : 1 During nonbreeding season, the giant panda′s ovary was still rich with large and small follicular oocytes, and the large follicular oocytes could be matured in vitro; 2 The mean diameter of the oocytes was (141±6.7)μm and lots of cortical granules contained in the cytoplasm of oo...

结果表明∶（1）在非繁殖季节，大熊猫卵巢中小腔及大腔卵泡卵母细胞仍较丰富，其大腔卵泡卵母细胞可在体外培养成熟；（2）大熊猫的卵母细胞平均直径为（141±6.7）μm ，其胞质富含卵黄颗粒；（3）自配 PandaI I号获能液（含丙酮酸钠、肾上腺素等）可使大熊猫精子有效获能；（4）大熊猫卵母细胞体外受精后，经培养可观察到第2 极体，有的受精卵开始发生卵裂。

The result of frozen MⅡoocytes using three methods indcated that the self-restraintspear method was better than OPS, and the effection of frozen altogether with self-restraintspear method and centrifugal was the best; the frozen effection was not ideal using fluidspear for 8- cell ovocyte.

用三种冷冻方法(OPS法；自制移液枪头法；移液枪头法+离心法)对MⅡ卵母细胞冷冻的结果表明，自制移液枪头法优于OPS法，其中以移液枪头法+离心组的冷冻效果最好；用移液枪头法对体外发育到8-细胞期卵母细胞的冷冻效果不理想。MⅡ期卵母细胞用OPS法冷冻后卵母细胞IVF和电激活的卵裂率都显著高于移液枪头法(P＜0.01)，电激活后2-细胞卵裂率比IVF高。

The cleavage rate of ovocyte IVF of MⅡovocyte frozen withOPS method and activation method was higher remarkably than fluid spearmethod(P＜0.01), the cleavage rate of 2-cell after electrical activation was higher than IVF.

本试验推荐猪体外成熟培养体系为连续培养液48h的NCSU-23+10％FBS，细胞浓度为100个／400μl，发育培养体系为NCSU-23+BSA；用CRY-3细胞激活仪电激活时(电融槽宽度为1mm)为150v／20μs／2次+600μMγ-BLI；以活力为0.1左右的冷冻精液进行IVF时，精子浓度为10~7，共孵时间为6h，并在成熟处理时只部分去除卵丘细胞；MⅡ卵母细胞冷冻时，方法为移液枪头法+离心法，冷冻后激活的2-细胞卵裂率比IVF高。

This experiment aimed to systematically investigate and analyze the ethanol activated rates of mouse oocytes, affected factors of oocytes activation, types and development of parthenotes, and the organization and roles of the cytoskeleton during pronuclear formation and migration and early embryo cleavage by ethanol activation of oocytes, embryo culture in vitro and immunofluorescence cyto-chemistry. The results:(1) The ethanol activation of mouse oocytes showed that the activated and developmental rates of oocytes increased significantly with the increase of ethanol concentration and extension of exposure time, but over concentration and exposure time would result in increased fragment rates significantly. 7% ethanol treated oocytes for 7min was the optimum activated condition.

本实验主要利用卵母细胞的乙醇激活、胚胎体外培养和免疫荧光细胞化学方法对小鼠卵母细胞乙醇激活效率、影响因素、孤雌胚类型、发育、原核形成、迁移及早期胚胎卵裂过程中细胞骨架的组装、作用等进行了系统的研究和分析，结果显示：(1)小鼠卵母细胞的乙醇激活结果表明，随着乙醇浓度的升高和作用时间的延长，卵母细胞激活率和发育率都显著提高，但乙醇浓度过高和作用时间过长会导致卵母细胞碎裂率的显著增加，7%乙醇作用卵母细胞7min 为最佳激活条件。

Our study includes four aspects. In the first aspect we study several important conditions of porcine oocytes maturation in vitro and oocytes cleavage after parthenogenetic activation and found mNCSU-23+15IU/mlPMSG+20IU/mlHCG+15% PFF+0.57mMcysteine is a good culture condition .When the Cocs are cultured in it ,the maturation rate and oocytes cleavage rate are higher than those of foreign covered. Our result are (86.7±3.35)% and (86.3±4.16)% and the highest report of foreign is(85.7±4.1)%.In the second aspect we study the effect of different chemical activations on development of porcine parthennogenetic embryo and found two best activation method. The first one is that putting the maturation MII oocytes in the 20μmol/L ionomycin for 30 minutes and then putting them in the NCSU-23 condition containing 5μg/mICB and 5mM/L6-DMAP for 3.5 hours, the oocytes cleavage rate and morulae/blastocysts development rate are (76.7±7.6)% and (37.1±6.4)%.The second one is that putting the maturation MII oocytes in the 200μM/L Thimerosal for 20 minutes and then putting them in the NCSU-23 condition containing 8mM DTT for 30 minutes

本研究分为4个部分，第一部分对影响猪卵母细胞体外成熟和孤雌激活后胚胎分裂的几个重要条件进行了比较研究，确立了一种较好的培养方法：与颗粒细胞共培养，找到了一种适合猪卵母细胞体外成熟的培养基：mNCSU-23+15IU/mlPMSG+20IU/mlHCG+15%PFF+0.57mM半胱氨酸，成熟率和分裂率分别为(86.7±3.35)%和(86.3±4.16)%，国外报道的最高成熟率为(85.7±4.1)%；第二部分对猪卵母细胞孤雌激活的化学方法进行了研究，确立了化学激活猪卵母细胞的两种最佳方法：1将成熟的去卵丘颗粒细胞的MII期卵母细胞用20μmol/Lionomycin作用30min，再将卵母细胞培养于含5μg/mlCB和5mM/L 6-DMAP(6-二甲基氨基嘌呤)的NCSU-23培养液中，卵裂率和桑囊胚发育率达到(76.7±7.6)%和(37.1±6.4)%2将成熟的去卵丘颗粒细胞的MII期卵母细胞在200μM/L的Thimerosal中处理20min，再与8mM的DTT共孵育30min，卵裂率和桑/囊胚形成率为(81.0±2.8)%和(39.6±2.7)%；第三部分对孤雌激活胚胎的培养条件进行了研究，确立了一种最佳的胚胎培养条件：在SOF简单培养基中添加颗粒细胞进行前3天的培养，然后转入添加胎牛血清的NCSU—23培养基并和输卵管上皮细胞进行后期的培养，其桑椹胚和囊胚的发育率为(59.5±3.2)%；第四部分研究了IGF-I

The sixth, there were no significantly differences between colchicine groupsand vinblastine groups on the metaphases, in the experiment of prepare mouse4-cell embryo stage single blastomere chromosome specimen by using optimumparameters; and there were significantly differences between colchicine groupsand vinblastine groups on the metaphases (P＜0.05), in the experiment of preparemouse 8-cell embryo stage single blastomere chromosome specimen by usingoptimum parameters.The end, the preparation quality of mouse 4-and 8-cell embryo stage singleblastomere specimen were 2.5, 2.7, 2.4, 1.9, respectively.

低渗液选用0.45％氯化钠时，低渗效果优于选用0.5％柠檬酸钠和0.35％氯化钾的。6、在利用最佳参数制备小鼠4-细胞期胚胎单卵裂球染色体标本时，经统计分析，两种阻断剂的作用效果差异不显著；而在制备小鼠8-细胞期胚胎单卵裂球染色体时阻断剂选用长春花碱的效果优于秋水仙素的(P＜0.05)。7、秋水仙素组和长春花碱组小鼠4、8-细胞胚胎单卵裂球染色体标本制备质量分别为：2.5、2.7、2.4、1.9。

On the other hand, the more important thing is because the urban housing is a kind of heterogeneity products.

另一方面，更重要的是由于城市住房是一种异质性产品。

Climate histogram is the fall that collects place measure calm value, cent serves as cross axle for a few equal interval, the area that the frequency that the value appears according to place is accumulated and becomes will be determined inside each interval, discharge the graph that rise with post, also be called histogram.

气候直方图是将所收集的降水量测定值，分为几个相等的区间作为横轴，并将各区间内所测定值依所出现的次数累积而成的面积，用柱子排起来的图形，也叫做柱状图。