Generation of tetraploid complementation mice from embryonic stem cells cultured with chemical defined medium  

作　　者：Chunjing Feng Haifeng Wan Xiao-Yang Zhao Liu Wang Qi Zhou 

机构地区：[1]State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences [2]University of Chinese Academy of Sciences [3]Key Laboratory of Translational Stem Cell Research, Institute of Zoology, Chinese Academy of Sciences

出　　处：《Chinese Science Bulletin》2014年第22期2743-2748,共6页

基　　金：supported by the National Basic Research Program of China(2012CBA01300 and2012CB966500)

摘　　要：Mouse embryonic stem cells(mESCs)derived from inner cell mass(ICM)of pre-implantation embryos,can maintain undifferentiated state when cultured in N2B27 medium supplemented with GSK3inhibitor CHIR99021 and MEK inhibitor PD0325901(‘‘2i’’)and leukemia inhibitor factor(LIF).Compare to conventional culture medium,all components of this medium are defined.With the N2B27 medium,‘‘2i’’and LIF,mESCs can contribute to the germline of the chimeric embryos,however,whether the‘‘all-ES cells’’mice can been generated by tetraploid complementation is unclear yet,while the tetraploid complementation serve as a golden standard to assess the pluripotency of ES cells.Here,our study showed that mESCs derived and cultured with the N2B27 complete medium could generate fertile mice by tetraploid complementation.In addition,the survival rate of tetraploid complementation mice produced by inbred mES cell lines is higher than the conventional culture condition,and increased the percentage of Oct4 positive cells contrast to conventional medium either.Therefore,the N2B27 medium supplemented with‘‘2i’’and LIF is an alternative choice forthe derivation and long-term culture of mouse embryonic stem cells.Mouse embryonic stem cells （mESCs） derived from inner cell mass （ICM） of preimplantation embryos, can maintain undifferentiated state when cultured in N2B27 medium supplemented with GSK3 inhibitor CHIR99021 and MEK inhibitor PD0325901 （“2i”） and leukemia inhibitor factor （L1F）. Compare to conventional culture medium, all components of this medium are defined. With the N2B27 medium, “2i” and LIF, mESCs can contribute to the germline of the chimeric embryos, however, whether the “all-ES cells” mice can been generated by tetraploid complementation is unclear yet, while the tetraploid complementation serve as a golden standard to assess the pluripotency of ES cells. Here, our study showed that mESCs derived and cultured with the N2B27 complete medium could generate fertile mice by tetraploid complementation. In addition, the survival rate of tetraploid complementation mice produced by inbred mES cell lines is higher than the conventional culture condition, and increased the percentage of Oct4 positive cells contrast to conventional medium either. Therefore, the N2B27 medium supplemented with “2i” and LIF is an alternative choice for the derivation and longterm culture of mouse embryonic stem cells.

关 键 词：小鼠胚胎干细胞 培养基 四倍体 互补 白血病抑制因子 ES细胞系 化学 植入前胚胎 

分 类 号：Q813[生物学—生物工程]