猪去核无透明带卵母细胞电融合参数的探讨

Abstract

Abstract: In the present study, we investigated the effect of electrical fusion parameters on pig oocytes of enucleated zona-free by using different electrical field strength and the pulse duration and then observed cell fusion. The results showed that the comparison study of different electrical fusion parametes indicated that the rate of oocytes using one DC-pulse of 0.8 kV/cm for 30 μs (94.7%) groups were significantly higher than the other treatment groups(P<0.05). The rate of fusion of the enucleated zona-free oocytes in one DC-pulse of 0.8 kV/cm for 30, 60 and 90 μs group were 94.7%, 86.5% and 83.8%, respectively, showing that 30 μs treatment group were significant higher than those of 60 and 90 μs treatment groups(P<0.05). The rate of fusion in one, double and three DC-pulse of 0.8 kV/cm for 30 μs were 94.7%, 95.6% and 81.7%, respectively, showing that double DC-pulse treatment groups were no significant higher than those of one DC-pulse treatment groups （P>0.05）, but both were significant higher than three DC-pulse treatment groups, the fusion rate of them was significant higher than three DC-pulse treatment groups（P<0.05）.

Key words: zona-free oocytes; fusion; porcine

ZHANG Bin, HUA Zai-dong, ZHENG Xin-min, XU Sheng-cheng. Exploration on Electrical Fusion Parameters of Enucleated Zona-free Oocytes[J]. .

References

1 文端成, 毕春明, 朱子玉, 等.卵裂球电脉冲融合制作昆明小鼠四倍体胚胎［J］.动物学研究, 2003, 24(2): 81～85.
2 任子利, 赵彦玲, 杨小淦, 等.猪手工体细胞核移植电融合/激活参数的优化［J］.畜牧兽医学报, 2010, 41(10): 1246～1252.
3 任子利, 赵彦玲, 何若钢, 等.有无透明带猪卵细胞电激活参数的优化及体外培养方式的建立［J］.安徽农业科学, 2008, 36(33): 14541～14544.
4 陆凤花, 石德顺, 韦英明, 等.不同融合条件对水牛体细胞核移植效果的影响［J］.广西农业生物科学, 2005, 24(1):1～5.
5 李日聪, 王锦, 薛林涛, 等.猪体细胞核移植电融合参数的研究［J］.广西农业生物科学, 2008, 27（4）:304～308.
6 李光华, 叶绍辉. 动物体细胞克隆技术的研究进展［J］.家畜生态学报, 2006, 27（4）:1～5.
7 孟庆刚, 朱士恩, 曾申明, 等.减少细胞质对激活小鼠卵母细胞体外发育的影响［J］.中国农业科学, 2003, 36(9): 1109～1112.
8 杨利军, 李煜. 无透明带体细胞核移植研究进展［J］. 中国畜牧兽医, 2009, 36(3): 52～57.
9 周琪, 李子义, 谭景和, 等. 核质比对家兔胚胎发育的影响［J］.科学通报, 1999, 44(3): 289～291.
10 Berg H. Fusion of blastomeres and blastocysts of mouse embryos［J］. Bioelectrochem Bioenerg, 1982, 9:223～228.
11 Bordignon V, Smith L C. Telophase enucleation: An improved method to prepare recipient cytoplasts for use in bovine nuclear transfer［J］. Mol Reprod Dev, 1998, 49(1):29～36.
12 Campbell K H, McWhir J, Ritchie W A, et al. Sheep cloned by nuclear transfer from a cultured cell line［J］. Nature, 1996, 380:64～66.
13 French A J, Adams C A, Anderson L S, et al．Development of human cloned blastocysts following somatic cell nuclear transfer with adult fibroblasts［J］．Stem Cells, 2008, 26: 485～493.
14 Greising T, Monaghan P, Healy A, et al. The influence of cytoplasmic composition and quantity of cytoplasm in recipient oocytes on the developmental capacity of bovine nuclear transfer embryos［J］. Theriogenology, 1994, 41: 208.
15 Kubiak J Z, Tarkowski A K. Electrofusion of mouse blastomeres［J］. Exp Cell Res, 1985, 157: 561～566.
16 Mau J C, Grupen C G, Verma P J, et al. MPF activity of porcine oocytes following electrical activation with and without 6-DMAP［J］. Theriogenology, 2001, 55:457～459.
17 Northey D L, Nuttleman P R, Rosenkrans C F. Removal of bovine oocyte cytoplasm prior to fertilization reduces cell number in embryos［J］. Biology of Reproduction, 1991, 44: 156.
18 Onishi A, Iwamoto M, Akita T, et al. Pig cloning by microinjection of fetal fibroblast nuclei［J］. Science, 2000, 289（5482）:1188～1190.
19 Wilmut I, Schnieke A E, McWhir J, et al．Viable offspring derived from fetal and adult mammalian cells［J］．Nature, 1997, 385:810～813.
20 Zakhartcheonko V, Stojkvic M, Brem G, et al. Enucleation of bovine oocytes with minimal cytoplasmic volume: Effect on development of nuclear transfer embryos［J］. Theriogenologya, 1997a, 47 (1): 238.
21 Zakhartchenko V, Stojkovic M, Brem G, et al. Karyoplast-cytoplast volume ratio in bovine nuclear transfer embryos: Effect on developmental potential［J］. Mol Reprod Dev, 1997b, 48(3): 332～338.

Exploration on Electrical Fusion Parameters of Enucleated Zona-free Oocytes

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 0

Recommended Articles

Metrics

Comments