ICSI受精卵胞质注射生产转基因猪胚胎的初步研究

doi:10.16431/j.cnki.1671-7236.2018.06.024

Abstract

Abstract:

This study was aimed to explore the feasibility of using intracytoplasmic sperm injection (ICSI) method to produce porcine in vitro fertilization (IVF) eggs and application of porcine ICSI fertilized eggs for cytoplasmic injection to produce transgenic embryos.First of all,we compared the efficiency of embryo development between porcine IVF zygotes and ICSI zygotes,and then applied cudbear staining method to observe the pronuclear formation at different time interval and efficiency produced by porcine ICSI zygotes,finally we injected the EGFP-N1(20 ng/μL) plasmid into the zygotes by ICSI and IVF methods to observe the embryonic development efficiency and transgene efficiency.The results showed that the ICSI fertilized eggs had better embryo development rate (cleavage rate 89.4% and 67.9%,blastocyst rate 36.5% and 16.1%) than IVF group(P<0.05);The pronuclear formation time by ICSI method was 12 to 14 h after sperm injection into the cytoplasm,and the rate of prokaryotic nuclei formation was 54.90%,which was significantly higher than the other 5 experimental groups (P<0.05);There was significant difference in the rate of embryonic cleavage rate (86.2% and 66.3%),blastocyst rate (30.0% and 13.6%) and transgene efficiency (18.5% and 0) between ICSI cytoplasmic injection group and IVF test group (P<0.05).This study laid the foundation for the study of producing transgenic pigs by cytoplasmic injection with ICSI fertilized eggs.

Key words: ICSI; cytoplasmic injection; transgene; pig embryos

CLC Number:

S814.8

LI Hongli, SU Xiaoping, LU Suyu, LIU Shuai, LIU Qingyou, CUI Kuiqing, SHI Deshun. Study on Transgenic Porcine Embryos by Cytoplasmic Injection into Zygotes Produced Using ICSI Method[J]. , 2018, 45(6): 1618-1625.

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Study on Transgenic Porcine Embryos by Cytoplasmic Injection into Zygotes Produced Using ICSI Method

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