ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2015, Vol. 46 ›› Issue (10): 1784-1790.doi: 10.11843/j.issn.0366-6964.2015.10.011

Previous Articles     Next Articles

Analysis of Factors Influencing Embryonic Development after Mouse or Pig Embryonic Stem Cells or Embryonic Germ(ES/EG) Mircoinjection

DONG Xiao1,FENG Shu-tang2* ,WANG Hong-jun1*   

  1. (1.College of Life Science,Qingdao Agricultural University,Qingdao 266109,China;2.Institute of Animal Science,Chinese Academy of Agricultural Sciences,Beijing 100193, China)
  • Received:2015-02-11 Online:2015-10-23 Published:2015-10-23

Abstract:

The goal of this study was to assess how different microinjection conditions influence chimeras production after ES/EG cells injection into embryos in mouse and pig.Mouse ES cells were micrioinjected into embryos at morula or blastocyst stages.Hatching rate and blastocyst rate after cell injections were measured in vitro.Non-injected embryos were used as controls.In mouse,embryos showed significant higher developmental rate when ES cells were injected into morulae compared to those injected into blastocysts(P<0.01).In contrast,cells injected into blastocysts had higher hatchability than those injected into morulae(P<0.01).Compared to non-injected controls,ES cell injected embryos had higher hatchability but lower developmental rate after in vitro culture(P<0.01).We further assessed porcine embryonic development after EG cells microinjection into morulae,early blastocysts,blastocysts,or hatched blastocysts,and we also investigated the optimal injecting/ holding pipette sizes for pig EG cell microinjection.Our data showed that pregnancy rates of EG cells microinjected into morulae,early blastocysts,or blastocysts,were significant higher than those injected into the hatched blastocysts(P<0.01).The hatched rate was significantly lower in microinjected embryos compared to non-injected controls(P<0.01).Two chimeric piglets were obtained after embryo transfer.In addition,we found that the optimal inner and outside diameters for pigs EG microinjection were 25 and 30 μm for the injecting pipette,and 40 μm and 130-150 μm for the holding pipette,respectively.Upon comparison of different conditions,we determined optimal injecting/holding pipette sizes for pig EG cell injection.Furthermore,ES/EG cells microinjection into morulae could achieve higher contribution rate in chimera.Microinjection had lower impact on mouse embryonic development compared to pig,suggesting that pig embryonic microinjection condition has to be further optimized.

CLC Number: