亮甲酚蓝染色对牛卵母细胞体外成熟的影响

›› 2009, Vol. 36 ›› Issue (2): 82-85.

亮甲酚蓝染色对牛卵母细胞体外成熟的影响

乔利敏1,2，黄成定2，李向臣1，关伟军1，马月辉1

(1.中国农业科学院北京畜牧兽医研究所，北京 100193； 2.甘肃农业大学动物科技学院，兰州 730070)

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2009-02-20 发布日期:2009-02-20
通讯作者: 关伟军

Effect of BCB Staining on the IVM of Cow Oocytes

QIAO Limin1,2, HUANG Chengding2, LI Xiangchen1, GUAN Weijun1, MA Yuehui1

(1.Institute of Animal Science(IAS),Chinese Academy of Agricultural Science(CAAS),Beijing 100193,China;2.College of Animal Science and Technology, Gansu Agriculture University, Lanzhou 730070,China)

Received:1900-01-01 Revised:1900-01-01 Online:2009-02-20 Published:2009-02-20
Contact: GUAN Wei-jun

摘要/Abstract

摘要： 为了探讨不同浓度亮甲酚蓝对牛卵母细胞染色选择后对其体外成熟（IVM）的影响，本研究在牛卵母细胞成熟培养前直接用不同浓度梯度的亮甲酚蓝进行不同时间的染色。结果表明：①经过不同浓度的亮甲酚蓝染色后，所选择卵母细胞染色后着色的百分率及其成熟率是不同的。用26、39、52 μmol/L的亮甲酚蓝染色后所得到的胞质蓝色组（27.7%、27.6%、31.9%）较13 μmol/L的亮甲酚蓝染色后所得到的胞质蓝色组（2.1%）差异显著（P＜0.05）；②经过不同浓度的亮甲酚蓝染色后，胞质蓝色组（77.4%）较胞质无色组（51.3%）和对照组（60.2%）成熟率差异显著（P＜0.05）；③不同浓度亮甲酚蓝染色的胞质蓝色组间成熟率差异不显著（P＞0.05）；④不同浓度亮甲酚蓝染色组与对照组卵母细胞的成熟率差异不显著（P＞0.05）；⑤经过不同时间的亮甲酚蓝染色后，所选择卵母细胞着色的百分率是不同的。用26 μmol/L的亮甲酚蓝染色90 min后所得到的胞质蓝色组（75%）较亮甲酚蓝染色60、30 min后所得到的胞质蓝色组（57.14%，40%）差异显著（P＜0.05）。以亮甲酚蓝染色为基础的牛卵丘卵母细胞复合体的区分可以用来有效的选择更具发育活力的牛卵母细胞。

关键词: 牛; 卵母细胞; 体外成熟; 亮甲酚蓝染色; 浓度; 葡萄糖-6-磷酸脱氢酶

Abstract: The aim of the present investigation was to study the effect of IVM of cow with different concentration brilliant cresyl blue (BCB) staining on the oocytes selection. For this purpose, Compact cumulus oocyte complexes (COCs) were recovered from slaughterhousecollected bovine ovaries and classified as control group, which were cultured immediately without exposure to BCB stain;treatment group were stained with different concentrations BCB(13,26,39 and 52 μmol/L BCB) for different time before culture. Treated oocytes were then classified according to their cytoplasm coloration: oocytes with a blue cytoplasm or grown oocytes(BCB+) and oocytes without a blue cytoplasm or growing oocytes(BCB-) . Oocytes were stained with different concentration BCB for different time before culture in this study. The result showed that:①The percentage of Colouring and mature for selected oocytes was different by treated with different concentrations BCB. The percentages of BCB+ oocytes obtained after staining with 26 μmol/L (27.7%),39 μmol/L (27.6%)and 52 μmol/L BCB (31.9%) were higher (P<0.05) than those obtained with 13 μmol/L(2.1%); ②BCB+ oocytes exhibited a higher (P<0.05) maturation rate (mean rate:77.4%;n=62)than BCB(51.3%;n=236) and control oocytes (60.2%;n=98);③There were no significant differences in the percentages of BCB+ oocytes reaching the MII stage among BCB concentrations;④There were no significant differences in nuclear maturation between control and BCB exposed oocytes (BCB+ plus BCB- ),independent of the BCB concentration;⑤After exposure different time BCB, the result showed that the percentage of BCB+ oocytes were different. The percentages of BCB+ oocytes obtained after staining with 26 μmol/L BCB for 90 min(75%) were a higher(P<0.05)than those oocytes obtained after staining with 26 μmol/L BCB for 60 min(57.14%) and 30 min(40%). These results show that the staining of cow cumulusoocyte complexes with BCB before in vitro maturation could be used to select developmentally competent oocytes.

Key words: cow; oocytes; IVM; brilliant cresyl blue; concentration; G6PDH

中图分类号:

S823.3

乔利敏;;黄成定;李向臣;关伟军;马月辉. 亮甲酚蓝染色对牛卵母细胞体外成熟的影响[J]. , 2009, 36(2): 82-85.

QIAO Limin;;HUANG Chengding;LI Xiangchen;GUAN Weijun; MA Yuehui . Effect of BCB Staining on the IVM of Cow Oocytes[J]. , 2009, 36(2): 82-85.

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