四倍体胚胎发育阶段对胚胎干细胞嵌合体小鼠制备的影响  

Effects of developmental stages of tetraploid embryos on mouse chimeras from embryonic stem cells

作　　者：吴兴龙[1] 张鹏[1] 赵虎[1] 王鹏博[1] 胡春超[1] 李相运[1] 

机构地区：[1]河北农业大学动物科技学院,河北保定071001

出　　处：《解剖学报》2011年第5期690-694,共5页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(30871790);河北省自然科学基金资助项目(C2009000600)

摘　　要：目的探讨四倍体胚胎发育阶段对胚胎干细胞(ES)嵌合体小鼠制备的影响。方法通过2-细胞胚胎电融合法制备四倍体胚胎,采用显微注射方法将ES细胞分别注入1-细胞、4-细胞、囊胚3个发育阶段的四倍体胚胎中。所用ES细胞分别为杂交系B6D2F1×129/Sv和近交系C57BL/6J,经胚胎移植和剖腹产以获得ES小鼠。结果实验表明,2-细胞胚胎电融合率为92.45%,4-细胞胚胎发育率为93.51%,囊胚发育率为90.42%。杂交ES细胞注射四倍体囊胚获得22只ES小鼠,效率显著高于近交系ES细胞以及其他发育阶段的四倍体胚胎,ES小鼠与B6D2F1×129/Sv杂交小鼠毛色一致且具有正常生殖能力。结论四倍体胚胎的发育阶段显著影响ES小鼠的制备。Objective To investigate the effects of developmental stages of tetraploid embryos on mouse chimeras from embryonic stem cells （ES）. Methods Tetraploid embryo complementation and microinjection were utilized to produce ES mice that derived completely from ES cells. Tetraploid embryos were firstly prepared by electrofusion of 2-cell mouse embryos, and then embryonic stem （ES） cells with different genetic background （hybrid or inbred） were injected into tetraploid 1-cell, 4-cell, and blastocyst stage embryos. The injected embryos were transferred into uterine horns of pseudopregnant 2.5days female CD1 mice. The CD1 mice were subjected into cesarean sections after 16 days. Results Our data showed that 92.45% of 2-cell embryos were electrofused, 93.51% and 90. 42% of the electrofused embryos developed to 4-cell and blastocyst stages, respectively. Blastocysts were injected with hybrid ES cells and 22 ES mice were obtained by Cesarean section. Blastoeyst was more effective than 1-cell and 4-cell. ES mice had normal germline transmission capability and the same coat color as the hybrid mice which hybrid ES cells were drived from. Conclusion The tetraploid embryo developmental stage is able to affect the ES mouse generation.

关 键 词：胚胎干细胞 四倍体胚胎 电融合 显微注射 嵌合体小鼠 

分 类 号：Q132.8[生物学—普通生物学]