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| Study on Generation of Chimeric Porcine Embryos Derived from Lentivirus Mediated Green Fluorescent Protein(GFP) Transgene Embryos |
| SHI Yong-qian, HE Wen-teng, ZHOU Yang, JIAO Ming-xia, XIE Bing-teng, HU Kui, KONG Qing-ran, LIU Zhong-hua |
| North-east Agricultural University, Harbin 150030, China |
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Abstract Objective: Through the establishment system of the lentiviral vector infected porcine embryo, studied chimeric ability of porcine parthenogenetic embryos of the different development stages, made a foundation for further studying early embryonic development and cell differentiation. Method: The lentivirus with GFP report gene was injected into the perivitelline space of porcine embryos at 1-cell and 2-cell stage, and the cleavage rate, GFP-positive embryo rate, blastocyst rate, GFP-positive blastocyst rate and cell number of blastocyst were examined in fluorescence microscope. Well of well (WOW) method was employed to make chimeric embryos by aggregating two different embryos at the same developmental stage of 2-cell, 4-cell and 8-cell or 2-cell stage embryo with 2-cell, 4-cell and 8-cell stage embryos, and blastomere exchanging at 2-cell stage was also used for making chimeric embryo.Then the chimerism of porcine embryos were checked. Result: In groups of 2?109I.U./ml lentivirous injecteion in the perivitelline of the 2-cell stage embryos, the infection rate and GFP-positive blastocyst rate of IVF(80.00%, 90.74%) and parthenogenetic embryos (76.36%, 89.56%) were significantly higher than other groups (P<0.05), and there is no significant difference in cleavage rate, blastocysts rate, blastocysts cell number compared with the control group. The chimeric rate of embryos derived from aggregating of two 2-cell stage embryos (53.85%) and embryos from blastomere exchanging of 2-cell stage embryos (62.50%) were significantly higher than that of aggregating of 2-cell with 4-cell or 8-cell embryos (18.60%, P<0.05), and the chimeric rate of embryos derived from aggregating of two 8-cell embryos (75.00%) were significantly higher than that of these embryos derived from aggregating of two 4-cell embryos (65.00%)or 2-cell embryos (53.80%). Conclusion: 2?109I.U./ml of lentivirous injection in the perivitelline of two-cell stage embryos shows the highest infection efficiency,in addition,lentivirus infection has no significant effect on developmental of porcine embryos. The chimeric rate of embryos derived from aggregating two 8-cell embryos is higher than the others.The chimerism rate of the synchronized developmental embryos is higher than that of the non-synchronized embryos.
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Received: 14 March 2012
Published: 25 August 2012
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