Figure 1.Generation of NOD-scid II2rg^-1-extended pluripotent stem cells.(A)Schematic of two approaches used for generating NOD-scid II2rg^-1-extended pluripotent stem cells:de novo derivation from blastocysts(upper p...Figure 1.Generation of NOD-scid II2rg^-1-extended pluripotent stem cells.(A)Schematic of two approaches used for generating NOD-scid II2rg^-1-extended pluripotent stem cells:de novo derivation from blastocysts(upper panels)and chemical reprogramming from embryonic fibroblasts(lower panels).(B)Phase-contrast images of de novo derived outgrowth and EPS colonies for 17 passages in LCDM medium.Scale bars,100 pm.(C)qRT-PCR analysis of XEN marker genes expression during the chemical induction process(day 16).Error bars indicate SEM(n=2).(D)Co-immunostaining of XEN marker genes during the chemical induction process(day 16).Upper panels:GATA6 and SALL4;lower panels:SOX17 and SALL4.Scale bars,100 pm.展开更多
Recently we have established a new culture condition enabling the derivation of extended pluripotent stem(EPS)cells,which,compared to conventional pluripotent stem cells,possess superior developmental potential and ge...Recently we have established a new culture condition enabling the derivation of extended pluripotent stem(EPS)cells,which,compared to conventional pluripotent stem cells,possess superior developmental potential and germline competence.However,it remains unclear whether this condition permits derivation of EPS cells from mouse strains that are refractory or non-permissive to pluripotent cell establishment.Here,we show that EPS cells can be robustly generated from non-permissive NOD-sc/d Il2rg 1 mice through de novo derivation from blastocysts.Furthermore,these cells can also be efficiently generated by chemical reprogramming from embryonic NOD-sc/d II2rg-/-fibroblasts.NOD-sc/d II2rg-/-EPS cells can be expanded for more than 20 passages with genomic stability and can be genetically modified through gene targeting.Notably,these cells contribute to both embryonic and extraembryonic lineages in vivo.More importantly,they can produce chimeras and integrate into the E13.5 genital ridge.Our study demonstrates the feasibility of generating EPS cells from refractory mouse strains,which could potentially be a general strategy for deriving mouse pluripotent cells.The generation of NOD-sc/d II2rg-/-Yaqin Du and Ting Wang contributed equally to this work.Electronic supplementary material The online version of this article(https://doi.org/10.1007/s13238-018-0558-z)contains supplementary material,which is available to authorized users.EPS cell lines permits sophisticated genetic modification in NOD-scid II2rg-/-mice,which may greatly advance the optimization of humanized mouse models for biomedical applications.展开更多
文摘Figure 1.Generation of NOD-scid II2rg^-1-extended pluripotent stem cells.(A)Schematic of two approaches used for generating NOD-scid II2rg^-1-extended pluripotent stem cells:de novo derivation from blastocysts(upper panels)and chemical reprogramming from embryonic fibroblasts(lower panels).(B)Phase-contrast images of de novo derived outgrowth and EPS colonies for 17 passages in LCDM medium.Scale bars,100 pm.(C)qRT-PCR analysis of XEN marker genes expression during the chemical induction process(day 16).Error bars indicate SEM(n=2).(D)Co-immunostaining of XEN marker genes during the chemical induction process(day 16).Upper panels:GATA6 and SALL4;lower panels:SOX17 and SALL4.Scale bars,100 pm.
基金the National Key Research and Development Program of China(2016YFA01001002017YFA0103000)+4 种基金the National Natural Science Foundation of China(Grant Nos.31730059 and 31521004)the Guangdong Innovative and En trepreneurial Research Team Program(2014ZT05S216)the Science and Technology Planning Project of Guangdong Province,China(2014B020226001 and 2016B030232001)the Science and Technology Program of Guangzhou,China(201508020001)National Natural Science Foundation of China(Grant No.31571052).
文摘Recently we have established a new culture condition enabling the derivation of extended pluripotent stem(EPS)cells,which,compared to conventional pluripotent stem cells,possess superior developmental potential and germline competence.However,it remains unclear whether this condition permits derivation of EPS cells from mouse strains that are refractory or non-permissive to pluripotent cell establishment.Here,we show that EPS cells can be robustly generated from non-permissive NOD-sc/d Il2rg 1 mice through de novo derivation from blastocysts.Furthermore,these cells can also be efficiently generated by chemical reprogramming from embryonic NOD-sc/d II2rg-/-fibroblasts.NOD-sc/d II2rg-/-EPS cells can be expanded for more than 20 passages with genomic stability and can be genetically modified through gene targeting.Notably,these cells contribute to both embryonic and extraembryonic lineages in vivo.More importantly,they can produce chimeras and integrate into the E13.5 genital ridge.Our study demonstrates the feasibility of generating EPS cells from refractory mouse strains,which could potentially be a general strategy for deriving mouse pluripotent cells.The generation of NOD-sc/d II2rg-/-Yaqin Du and Ting Wang contributed equally to this work.Electronic supplementary material The online version of this article(https://doi.org/10.1007/s13238-018-0558-z)contains supplementary material,which is available to authorized users.EPS cell lines permits sophisticated genetic modification in NOD-scid II2rg-/-mice,which may greatly advance the optimization of humanized mouse models for biomedical applications.