| This study did microinjection to Kunming white mouse inbred embryos to test its in vitro developmental ability, then pluripotency of porcine EG cells was tested with production of chimera by blastocyst injection. Finally, first chimeric pig in China was delivered, and by chimera analysis, chimerism is present in some tissues. Experiment IStudy on the embryonic development by injecting different developmental stage embryos. This experiment collected 109 embryos to observe their developmental ability by ES cells microinjection into 8-16 cell stage morula or blastocyst. The result is that developmental rate and hatching rate of 57 microinjected 8-16 cell stage morulae is 94.7%(54/57)and 70.2%(40/57) separately, and that of 52 microinjected blastocysts is 86.4%(44/52) and 60.8%(42/52). The experiment shows that developmental rate and hatching rate between microinjected morulae and blastcysts has insignificant difference(P > 0.05). Technically, morula microinjection is easier. Experiment IIStudy on the production of porcine chimera by injecting embryonic stem cells (EG). The method of chimeric production is established in this experiment. External diameter of holding pipette is 130~ 150um, and internal diameter of injection pipette is 25um. hi this experiment, 311 embryos was obtained. In vitro cultivation developmental rate of 26 microinjected embryos is 65.4%(17/26). 164 microinjected embryos and 121 noninjected embryos were transferred to 14 recipient gilts.Of this, 159 microinjected and noninjected embryos(blastocyst, early blastocyst and morula) was transferred to 6 recipient gilts, three pigs are pregnant(one gilt delivered 8 piglets, two are skin pigment chimera), and pregnancy and chimera rate is 17%(l/6). 126 microinjected and nonmicroinjected expanded blastocysts oestrused on time, pregnancy and chimera rate is 0(0/5). Two stage embryos were observed their effect on production of chimera. The result shows that there is a significant different(0.01 |
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