梅花鹿-牛异种体细胞核移植操作方法的研究

doi:10.11843/j.issn.0366-6964.2014.08.026

Abstract

Abstract:

The purpose of this study was to optimize the manipulation of interspecies somatic cell nuclear transfer (ISCNT) between Sika deer and bovine.It was used the Sika deer ear skin fibroblast as nuclear donor cell，and bovine MII oocyte as recipient.McGrath-Solter，extrusion and DEME chemically assisted methods were used in enucleation.perivitelline microinjection (PM)，break-membrane-cell intracytoplasmic microinjection (BMCIM) and whole-cell intracytoplasmic microinjection (WCIM) were used in nuclear injection.Oocytes were electrofusion after enucleation of PM before chemical activated and others were chemically activated without electrofusion.Reconstructed embryos were cultured in media of CR1aa.The results showed that：(1) The rate of enucleation with the DEME method was the highest，and the rate of extrusion method was the lowest.There was significantly difference (P<0.05) in the rate of enucleation among all the three enucleate methods，but not significantly difference (P>0.05) in duration of enucleation.(2) The rate of nuclear injection by PM and WCIM was 100.0% and 94.8% separately，and the duration of nuclear injection by PM and WCIM was 31.0 and 35.1 s separately.There was no difference between them (P>0.05)，their rate of nuclear injection were higher than which in BMCIM group (P<0.05)，but duration of nuclear injection was converse (P>0.05).(3) The rate of activation by BMCIM and WCIM group were higher than it in PM group (P<0.05)，and which in BMCIM group of one-step method was the highest.There was no difference among the rate of activation，cleavage and blastocyst between one-step and two-step method (P>0.05).The results suggested that one-step manipulative method of DEME assisted enucleation and pizeo assisted BMCIM was efficient in Sika deer-bovine ISCNT.

CLC Number:

S814.8

WANG Shi-yong，ZHENG Jun-jun，YANG Yue-chun，LIU Zong-yue，YU Miao，YANG Fu-he. Study on Manipulation of Sika Deer-Bovine Interspecies Somatic Cell Nuclear Transfer[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(8): 1381-1386.

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Study on Manipulation of Sika Deer-Bovine Interspecies Somatic Cell Nuclear Transfer

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