Generation and characterization of stable pig pregastrulation epiblast stem cell lines.
Generation and characterization of stable pig pregastrulation epiblast stem cell lines.
复制标题
稳定猪原肠胚形成前外胚层干细胞系的产生和表征。
DOI：
10.1038/s41422-021-00592-9
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发表时间：
2022-04
期刊：
影响因子：
44.1
通讯作者：
Han J
中科院分区：
文献类型：
--
作者：
Zhi M;Zhang J;Tang Q;Yu D;Gao S;Gao D;Liu P;Guo J;Hai T;Gao J;Cao S;Zhao Z;Li C;Weng X;He M;Chen T;Wang Y;Long K;Jiao D;Li G;Zhang J;Liu Y;Lin Y;Pang D;Zhu Q;Chen N;Huang J;Chen X;Yao Y;Yang J;Xie Z;Huang X;Liu M;Zhang R;Li Q;Miao Y;Tian J;Huang X;Ouyang H;Liu B;Xie W;Zhou Q;Wei H;Liu Z;Zheng C;Li M;Han J
猪外胚层来源的多能干细胞被认为在人类治疗模型开发和家畜育种方面具有巨大的潜力和广阔的前景。尽管自20世纪90年代以来一直在进行尝试，但尚未建立稳定定义的猪外胚层衍生干细胞系。在这里，从大规模的单细胞转录组分析猪胚胎从胚胎天（E）0到E14的见解，特别是，在上胚层发育过程中的多能性变化的跟踪指导下，我们开发了一种体外培养基，用于建立和维持稳定的多能干细胞系从猪E10前原肠胚形成上胚层（pgEpiSC）。通过化学抑制WNT相关信号传导与FGF/ERK、JAK/STAT 3和激活素/Nodal途径中的生长因子的组合，pgEpiSC保持其多能性转录组特征，类似于E10上胚层细胞的转录组特征，并且在超过240代后保持正常核型，并且具有分化成三个胚层的潜力。引人注目的是，超深原位Hi-C分析揭示了染色质3D空间关联对pgEpiSC中多能性标记基因的转录调控的功能影响。在实践中，我们证实pgEpiSC容易耐受至少三轮连续的基因编辑并产生克隆的基因编辑的活仔猪。我们的发现兑现了人们期待已久的猪多能干细胞的承诺，并为生物研究、畜牧业和再生生物医学开辟了新的途径。
Pig epiblast-derived pluripotent stem cells are considered to have great potential and broad prospects for human therapeutic model development and livestock breeding. Despite ongoing attempts since the 1990s, no stably defined pig epiblast-derived stem cell line has been established. Here, guided by insights from a large-scale single-cell transcriptome analysis of pig embryos from embryonic day (E) 0 to E14, specifically, the tracing of pluripotency changes during epiblast development, we developed an in vitro culture medium for establishing and maintaining stable pluripotent stem cell lines from pig E10 pregastrulation epiblasts (pgEpiSCs). Enabled by chemical inhibition of WNT-related signaling in combination with growth factors in the FGF/ERK, JAK/STAT3, and Activin/Nodal pathways, pgEpiSCs maintain their pluripotency transcriptome features, similar to those of E10 epiblast cells, and normal karyotypes after more than 240 passages and have the potential to differentiate into three germ layers. Strikingly, ultradeep in situ Hi-C analysis revealed functional impacts of chromatin 3D-spatial associations on the transcriptional regulation of pluripotency marker genes in pgEpiSCs. In practice, we confirmed that pgEpiSCs readily tolerate at least three rounds of successive gene editing and generated cloned gene-edited live piglets. Our findings deliver on the long-anticipated promise of pig pluripotent stem cells and open new avenues for biological research, animal husbandry, and regenerative biomedicine.
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影响因子：
21.3
作者：
Boroviak T;Loos R;Bertone P;Smith A;Nichols J
通讯作者：
Nichols J
影响因子：
4.4
作者：
Cao S;Han J;Wu J;Li Q;Liu S;Zhang W;Pei Y;Ruan X;Liu Z;Wang X;Lim B;Li N
通讯作者：
Li N
影响因子：
14.9
作者：
Khan A;Fornes O;Stigliani A;Gheorghe M;Castro-Mondragon JA;van der Lee R;Bessy A;Chèneby J;Kulkarni SR;Tan G;Baranasic D;Arenillas DJ;Sandelin A;Vandepoele K;Lenhard B;Ballester B;Wasserman WW;Parcy F;Mathelier A
通讯作者：
Mathelier A
影响因子：
64.8
作者：
Dixon JR;Jung I;Selvaraj S;Shen Y;Antosiewicz-Bourget JE;Lee AY;Ye Z;Kim A;Rajagopal N;Xie W;Diao Y;Liang J;Zhao H;Lobanenkov VV;Ecker JR;Thomson JA;Ren B
通讯作者：
Ren B
影响因子：
64.8
作者：
Jin, Fulai;Li, Yan;Dixon, Jesse R.;Selvaraj, Siddarth;Ye, Zhen;Lee, Ah Young;Yen, Chia-An;Schmitt, Anthony D.;Espinoza, Celso A.;Ren, Bing
通讯作者：
Ren, Bing