BCB和Zebularine对绵羊体细胞核移植胚胎发育效果的影响

发布时间：2018-11-03 17:24

【摘要】：为探究亮甲酚蓝(BCB)染色选择的卵母细胞是否有利于体细胞核移植胚胎的体外发育,本实验将卵丘-卵母细胞复合体(COCs)放入含有BCB的PBS中染色,根据细胞质颜色可将卵母细胞分成BCB~+组和BCB~-组,并以未经BCB处理的COCs作为对照组,然后将卵母细胞进行体外成熟,统计卵母细胞的成熟率。将成熟后的卵母细胞进行体细胞核移植,其中,部分BCB~+组卵母细胞所需的供体细胞利用Zebularine处理,统计体细胞核移植的卵裂率、桑椹胚率和囊胚率。结果表明:BCB~+组卵母细胞的成熟率显著高于对照组和BCB~-组(71.15%vs 65.38%,53.52%,P0.05);BCB~+组核移植胚胎的卵裂率(87.91%vs 56.83%)、桑椹胚率(37.41%vs 21.73%)和囊胚率(21.48%vs6.82%)均显著高于BCB~-组(P0.05);与对照组相比,BCB~+组卵裂率(87.91%vs 83.23%)和囊胚率(21.48%vs14.89%)也显著升高(P0.05)。BCB~+组供体细胞经Zebularine处理后,胚胎发育能力进一步提高,其中囊胚率显著高于BCB~+组、对照组和BCB~-组(29.25%vs 21.48%,14.89%,6.82%,P0.05)。
[Abstract]:In order to investigate whether the oocytes selected by brilliant cresol blue (BCB) staining were beneficial to the development of somatic nuclear transfer embryos in vitro, the cumulus oocyte complex (COCs) was stained in PBS containing BCB. According to cytoplasmic color, oocytes were divided into BCB~ group and BCB~- group, and COCs without BCB treatment was used as control group. The oocytes were matured in vitro to calculate the maturation rate of oocytes. Somatic nuclear transfer was carried out on mature oocytes. The rate of cleavage, morula and blastocyst of somatic nuclear transfer was counted by Zebularine treatment of donor cells needed for some BCB~ oocytes. The results showed that the maturation rate of oocytes in BCB~ group was significantly higher than that in control group and BCB~- group (71.15%vs 65.38). The cleavage rate (87.91%vs 56.83%), morula rate (37.41%vs 21.73%) and blastocyst rate (21.48 vs 6.82%) in BCB~ group were significantly higher than those in BCB~- group (P0.05). Compared with the control group, the cleavage rate (87.91%vs 83.23%) and blastocyst rate (21.48 vs 14.89%) in BCB~ group were significantly higher than those in control group (P0.05). After treated with Zebularine, the embryonic development ability of donor cells in BCB~ group was further improved. The blastocyst rate was significantly higher than that in BCB~ group, control group and BCB~- group (29.25%vs 21.48 and 14.89%, 6.82%, P0.05).
【作者单位】：河北农业大学动物科技学院;河北省牛羊胚胎工程技术研究中心;
【基金】：河北省自然科学基金(C2014204119) 河北省高等学校科学技术研究项目(ZD2014002)
【分类号】：S826

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