ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2006, Vol. 37 ›› Issue (6): 560-565.doi:
• 遗传繁育 • Previous Articles Next Articles
YANG Wei-feng ;GE Xiu-guo;HUA Jin-lian;DOU Zhong-ying
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Abstract: Rabbit antigoatspleen and antigoatfibroblast serum were each produced in 2 New Zealand White rabbits separately which were bled 10 days after 3 intravenous injections of 4×108 cells into Guanzhong dairy goat spleen and fetal fibroblast cells. In each group 2 rabbits were immunized and the 4 kinds of antiserum were all cytotoxinic, but the lysis value of antiserum obtained from spleen was slightly higher than fetal fibroblast cells. 24 Guanzhong dairy goats and 3 Boer goats were superovulated and collected embryos during 6-9 d after mating with a result of total 240 embryos including 106 blastocysts. Our results show that the suitable embryo development time to separate inner cell mass (ICM) is 7 to 9 days after mating. Antiserum was cytotoxinic to goat red blood cells, embryos of Guanzhong dairy goats and Boer goats according to their cytotoxinic experiments. Comparing 50%dissolvable value of goat red blood cells with lysis trophoblast cell value, we find the latter is 22-23 times as compared with the former. Thus it is feasible to get the suitable lysis trophoblast cell value by immunosurgery treatment of red blood cells. Separation of ICM from goat blastocysts by immunosurgery is a twostep procedure. The best parameters were as follows: the proportion of antiserum dilution is 1∶4 to 1∶15, the proportion of complement dilution is 1∶20, the time for embryos exposed to antiserum is 30 min and to complement is about 20-30 min. Finally, the damaged trophoblastic layer was removed by pipetting the blastocysts through a small-bore glass pipette and the ICM could easily be separated from the remnants of trophoblastic cells. The morphology of ICM is closely related to the quality of its derived blastocyst. When ICM was seeded on mitomycin-inactivated mouse embryonic fibroblast feeders, they would attach on feed layers in 2 days. 4-7 days later, ICM proliferated and formed ES-like colonies which were surrounded by lots of vacuolar-like cells.
YANG Wei-feng;GE Xiu-guo;HUA Jin-lian;DOU Zhong-ying. Separation of Inner Cell Mass from Goat Blastocysts by Immunosurgery[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2006, 37(6): 560-565.
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