数字农科院2.0

Effects of different states of sheep fetal fibroblasts as donor cells on the early development in vitro of reconstructed sheep embryos

文献类型： 外文期刊

作者： Wang Hai;Ao Hong;Pan QiuZhen;Li RongQi;Zhao MengBin;Lian ZhengXing;Li Ning;Wu ChangXin

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关键词： sheep;nuclear transfer;colchicine;serum starvation;fetal fibroblast;GFP gene

期刊名称： SCIENCE IN CHINA SERIES C-LIFE SCIENCES

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收录情况： SCI

摘要： To investigate the effects of different states of donor cells on the development of reconstructed sheep embryos,we designed five treatments of donor cells,including cell passage,cell size,serum starvation,colchicine treatment and gene transfection.Results are as follows: (I) Compared with 16-18 passage cells,the morula/blastocyst rate of 5-7 passage cells as donor nuclei was significantly higher (17.3% vs.4.9%,P<0.05),suggesting the advantage of short-time cultured cells in supporting the development of reconstructed embryos.(II) The morula/blastocyst rate of reconstructed embryos derived from medium cells (15-25 mu m) as donor nuclei was higher than that from large cells (25-33 mu m) and small cells (8-15 mu m)( 20.0% vs.8.0%,9.7%),indicating that reconstructed embryos from medium cells had a greater potentiality to develop into morula/blastocysts than those from small or large ones.(IE) The morula/blastocyst rate of reconstructed embryos from donor cells of SS (serum starvation) was lower than that from donor cells of NSS (non-serum starvation),but no significant difference was detected between SS and NSS( 11.8% vs.18.6%,P>0.05).(IV) Fetal fibroblasts treated with 0.05 mu mol/L colchicine exhibited a higher morula/blastocyst rate of reconstructed embryos than those treated with 0.10 umol/L colchicine and untreated ones (27.5% vs.12.1%,17.1%),however,no significant difference among the three treatments was detected (P>0.05).(V) The morula/blastocyst rate of reconstructed embryos from fetal fibroblasts transfected with GFP gene only was 3.1%,significantly lower than that from non-transgenic cells (3.1% vs.20.4%,P<0.05).In conclusion,our results demonstrated that fetal fibroblasts of fewer passages,medium size could ensure a higher morula/blastocyst rate of reconstructed embryos.Serum starvation of donor cells might be unnecessary to the development of reconstructed embryos.Donor cells treated with 0.05 umol/L colchicine could facilitate the development of reconstructed embryos.Additionally,as cells transfected with GFP gene were used as donor nuclei,adverse effect on the development of reconstructed embryos was observed.Therefore,the developmental efficiency of reconstructed embryos could be improved if proper treatments to donor cells were used.

分类号： Q1

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