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Tetraploid embryos could be produced by electrofusion at the stage of two-cell embryos, which could develop to blastocysts followed by fusion of cytoplasm and nucleus and cleavage in vitro. During the fusion of cytoplasm, the DNA methylation levels of the fused embryos are as high as these of two-cell diploid embryos in vivo Then the embryos are rapidly demethylated when the nucleus begin to fuse, resulting in the lowest DNA methylation levels when the nucleus are fused completely. After that, the DNA methylation levels of the fused embryos are gradually increased until the morula stage. However, whereas an asymmetric distribution of DNA methylation is established in vivo-derived blastocysts with a higher methylation level in the inner cell mass than that in the trophectoderm, we can not detect the asymmetric distribution in most in vitro-derived tetraploid blastocysts.

结果表明:利用电融合方法制备的小鼠四倍体胚胎在体外培养体系中经历细胞质融合、细胞核融合及细胞继续分裂发育直到囊胚期的过程,在细胞质融合的时候胚胎卵裂球同体内体外培养二倍体胚胎一样,呈现高度甲基化状态;在细胞核开始融合的时候,甲基化水平急速下降,在细胞核完全融合的时候甲基化水平达到最低点;随着胚胎继续分裂,胚胎甲基化水平逐渐增加,在桑葚胚期甲基化水平最高;但是囊胚期四倍体胚胎内细胞团同滋养层细胞甲基化荧光信号没有差别,这与体内体外培养二倍体囊胚内细胞团细胞甲基化荧光强度高于滋养层细胞甲基化荧光强度不同。

Tetraploid embryos could be produced by electrofusion at the stage of two-cell embryos, which could develop to blastocysts followed by fusion of cyto-plasm and nucleus and cleavage in vitro. During the fusion of cytoplasm, the DNA methylation levels of the fused embryos are as high as these of two-cell diploid embryos in vivo Then the embryos are rapidly demethylated when the nucleus begin to fuse, resulting in the lowest DNA methylation levels when the nucleus are fused completely. After that, the DNA methy-lation levels of the fused embryos are gradually increased until the morula stage. However, whereas an asymmetric distribu-tion of DNA methylation is established in vivo-derived blastocysts with a higher methylation level in the inner cell mass than that in the trophectoderm, we can not detect the asymmetric distribution in most in vitro-derived tetraploid blastocysts.

结果表明:利用电融合方法制备的小鼠四倍体胚胎在体外培养体系中经历细胞质融合、细胞核融合及细胞继续分裂发育直到囊胚期的过程,在细胞质融合的时候胚胎卵裂球同体内体外培养二倍体胚胎一样,呈现高度甲基化状态;在细胞核开始融合的时候,甲基化水平急速下降,在细胞核完全融合的时候甲基化水平达到最低点;随着胚胎继续分裂,胚胎甲基化水平逐渐增加,在桑葚胚期甲基化水平最高;但是囊胚期四倍体胚胎内细胞团同滋养层细胞甲基化荧光信号没有差别,这与体内体外培养二倍体囊胚内细胞团细胞甲基化荧光强度高于滋养层细胞甲基化荧光强度不同。

Tetraploid embryos could be produced by electrofusion at the stage of two-cell embryos, which could develop to blastcysts fellowed by fusion of cytoplasm and nucleus and cleavage in vitro.After fusion of cytoplasm, the DNA methylation levels of the fused embryos was very high as well as two-cell diploid embryos in vivo.Then the embryos was rapiddly demethylated when the nucleus begin to fuse, resulting the lowest DNA methylation levels when the nucleus fused completely.After that, the DNA methylation levels of fused embryos were gradually increased until the blastocysts stage.However, whereas an asymmetric distribution of DNA methylation was established in an vivo-derived blastocysts with a higher methylation level in the inner cell mass than in the trophectoderm, in most vitro-derived tetraploid blastocysts, we can not detect the asymmetric distribution.

结果表明:利用电融合方法制备的小鼠四倍体胚胎在体外培养体系中经历细胞质融合、细胞核融合及细胞继续分裂发育直到囊胚期的过程,在细胞质融合的时候胚胎卵裂球同体内体外培养二倍体胚胎一样,呈现高度甲基化状态;在细胞核开始融合的时候,甲基化水平急速下降,在细胞核完全融合的时候甲基化水平达到最低点;随着胚胎继续分裂,胚胎甲基化水平逐渐增加,在囊胚期甲基化水平最高;但是囊胚期四倍体胚胎内细胞团同滋养层细胞甲基化荧光信号没有差别,这与体内体外培养二倍体囊胚内细胞团细胞甲基化荧光强度高于滋养层细胞甲基化荧光强度不同。

Trophoblast Medium is a complete medium designed for optimal growth of normal human trophoblast in vitro .

滋养层细胞培养基是专门为正常人类滋养层细胞体外培养设计的最适于其生长的培养基。

These results suggest that flow-induced shear stress regulates the expression of β1 integrin, in turn affects the motility of trophoblast cells.

流动剪切力的参与调节了滋养层细胞β1 整合素的表达,从而可影响滋养层细胞的迁移。

The methods of trophoblast isolation have deficiency of low product ion rate and other cells contamination.

原代培养的滋养层细胞存在产率低、杂细胞污染等方面的缺憾,而滋养层细胞又在细胞特性上发生了变化。

Uu not only bring out corresponding ultrapathological changes by adhering to the surface of the cytotrophoblast membrane,but also enters the body of the cytotrophoblast and does harm to them directly.

Uu不仅吸附在滋养层细胞膜表面诱发相应的超微病理改变,而且还进入滋养层细胞体内,对滋养层细胞产生直接杀伤作用。

From the early study of the pathogenesis of pre-eclampsia people have found its association with immunological function . In 1986 Ellis et al[1] had found that the extravillous chorionic cytotrophoblast contacting with the maternal tissue expressed high levels of Human leukocyte antigen-G, a major histocompatibility tissue-specific antigen . Extravillous chorionic cytotrophoblast with high levels expression of HLA-G proteins can invade the maternal deciduas and the maternal spiral arteries by displacing the endothelial lining and destroying most of the musculoelastic tissue of these arteries .

早期研究开始,许多学者即发现其与免疫学有密切关系,至1986年Ellis等[1]发现人类白细胞抗原(human leukocyte antigen,HLA) G分子高度表达在绒毛外滋养层细胞( extravillous cytotrophoblast,EVCT)上,即浸润至子宫蜕膜层甚至达子宫浅肌层以及子宫螺旋动脉内皮层,与母血及子宫组织直接接触的滋养细胞,这种独特的组织分布提示HLA-G分子在母胎免疫耐受方面起重要作用。

In the normal uterus, Cytokeratins immunolabelling were detected in glandular cell, luminal epithelial cell, Vimentin immunolabelling were detected in stromal cell and endoblastic cell; CK7 immunolabelling were not detected in any tissue of the yak utenus.

研究结果显示:未妊娠时,泛角蛋白在子宫内膜腺上皮细胞、腔上皮细胞内表达,波形蛋白在子宫内膜基质细胞内表达,平滑肌肌动蛋白在子宫平滑肌和血管平滑肌内表达,牦牛子宫任何部位均不表达角蛋白7;妊娠30天左右时,泛角蛋白在子宫内膜腺上皮细胞、子宫内膜腔上皮细胞、滋养层细胞、内胚层细胞和尿囊细胞内表达,波形蛋白在子宫内膜基质细胞和内胚层细胞内表达,平滑肌肌动蛋白在子宫平滑肌和血管平滑肌内表达,角蛋白7在尿囊细胞内表达,偶尔在腔上皮细胞的细胞核边缘表达;消化法进行原代培养时,组织经胶原酶消化并通过100目和400目筛网组合可以有效地分离原代子宫内膜基质细胞和子宫内膜腺上皮细胞;分离得到的子宫内膜基质细胞活率达90%以上,并可在体外传代7次以上;分离得到的子宫内膜腺上皮细胞活率可达85%以上,并可在体外传代5次以上;RPMI1640培养基最适合子宫内膜基质细胞和子宫内膜腺上皮细胞的生长,维持子宫内膜基质细胞正常生长的FBS添加量为20%,维持子宫内膜腺上皮细胞正常生长的胎牛血清添加量为30%。

The a -Gal remarkably positive staining was appeared in membrane and plasm on ES cell, but nuclear is negative. It indicates that we must pay attention to HAR in cell xenotransplantation, especially differentiation ES cell in vivo.

筛选过程中在选择性培养基中添加LIF和BMP一2替代滋养层细胞,发现ES细胞的生长没有受到影响,提示在做基因敲除胚胎干细胞的筛选中可以不需要制备Nco抗性的滋养层细胞。

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昨晚我没有看电视,因为电视机坏了。

Since this year, in a lot of villages of Beijing, TV of elevator liquid crystal was removed.

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