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Generation and characterization of stable pig pregastrulation epiblast stem cell lines.
Zhi, Minglei; Zhang, Jinying; Tang, Qianzi; Yu, Dawei; Gao, Shuai; Gao, Dengfeng; Liu, Pengliang; Guo, Jianxiong; Hai, Tang; Gao, Jie; Cao, Suying; Zhao, Zimo; Li, Chongyang; Weng, Xiaogang; He, Mengnan; Chen, Tianzhi; Wang, Yingjie; Long, Keren; Jiao, Deling; Li, Guanglei; Zhang, Jiaman; Liu, Yan; Lin, Yu; Pang, Daxin; Zhu, Qianqian; Chen, Naixin; Huang, Jingjing; Chen, Xinze; Yao, Yixuan; Yang, Jingcang; Xie, Zicong; Huang, Xianya; Liu, Mengxin; Zhang, Ran; Li, Qiuyan; Miao, Yiliang; Tian, Jianhui; Huang, Xingxu; Ouyang, Hongsheng; Liu, Bofeng; Xie, Wei; Zhou, Qi; Wei, Hongjiang; Liu, Zhonghua; Zheng, Caihong; Li, Mingzhou; Han, Jianyong.
Afiliação
  • Zhi M; State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
  • Zhang J; State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
  • Tang Q; Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu, Sichuan, China.
  • Yu D; State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
  • Gao S; Institute for Stem Cell and Regenerative Medicine, Chinese Academy of Sciences, Beijing, China.
  • Gao D; Beijing Institute for Stem Cell and Regenerative Medicine, Beijing, China.
  • Liu P; Key Laboratory of Animal Genetics, College of Animal Science and Technology, China Agricultural University, Beijing, China.
  • Guo J; State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
  • Hai T; Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu, Sichuan, China.
  • Gao J; State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan Agricultural University, Kunming, Yunnan, China.
  • Cao S; State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
  • Zhao Z; State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
  • Li C; Animal Science and Technology College, Beijing University of Agriculture, Beijing, China.
  • Weng X; State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
  • He M; State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
  • Chen T; Key Laboratory of Animal Cellular and Genetics Engineering of Heilongjiang Province, College of Life Science, Northeast Agricultural University, Harbin, Heilongjiang, China.
  • Wang Y; Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu, Sichuan, China.
  • Long K; State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
  • Jiao D; State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
  • Li G; Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu, Sichuan, China.
  • Zhang J; State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan Agricultural University, Kunming, Yunnan, China.
  • Liu Y; School of Life Science and Technology, ShanghaiTech University, Shanghai, China.
  • Lin Y; Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu, Sichuan, China.
  • Pang D; Key Laboratory of Animal Cellular and Genetics Engineering of Heilongjiang Province, College of Life Science, Northeast Agricultural University, Harbin, Heilongjiang, China.
  • Zhu Q; Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu, Sichuan, China.
  • Chen N; Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Sciences, Jilin University, Changchun, Jilin, China.
  • Huang J; State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
  • Chen X; State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
  • Yao Y; State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
  • Yang J; State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
  • Xie Z; State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
  • Huang X; Animal Science and Technology College, Beijing University of Agriculture, Beijing, China.
  • Liu M; Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Sciences, Jilin University, Changchun, Jilin, China.
  • Zhang R; Animal Science and Technology College, Beijing University of Agriculture, Beijing, China.
  • Li Q; Animal Science and Technology College, Beijing University of Agriculture, Beijing, China.
  • Miao Y; State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
  • Tian J; State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China.
  • Huang X; Institute of Stem Cell and Regenerative Biology, College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei, China.
  • Ouyang H; Key Laboratory of Animal Genetics, College of Animal Science and Technology, China Agricultural University, Beijing, China.
  • Liu B; School of Life Science and Technology, ShanghaiTech University, Shanghai, China.
  • Xie W; Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Sciences, Jilin University, Changchun, Jilin, China.
  • Zhou Q; Center for Stem Cell Biology and Regenerative Medicine, MOE Key Laboratory of Bioinformatics, THU-PKU Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing, China.
  • Wei H; Center for Stem Cell Biology and Regenerative Medicine, MOE Key Laboratory of Bioinformatics, THU-PKU Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing, China.
  • Liu Z; State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
  • Zheng C; State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan Agricultural University, Kunming, Yunnan, China.
  • Li M; Key Laboratory of Animal Cellular and Genetics Engineering of Heilongjiang Province, College of Life Science, Northeast Agricultural University, Harbin, Heilongjiang, China. liuzhonghua@neau.edu.cn.
  • Han J; Key Laboratory of Genomic and Precision Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences, and China National Center for Bioinformation, Beijing, China. ch_zheng@aliyun.com.
Cell Res ; 32(4): 383-400, 2022 04.
Article em En | MEDLINE | ID: mdl-34848870
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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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Células-Tronco Pluripotentes / Camadas Germinativas Idioma: En Ano de publicação: 2022 Tipo de documento: Article
Texto completo
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XML
PubMed Links
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Células-Tronco Pluripotentes / Camadas Germinativas Idioma: En Ano de publicação: 2022 Tipo de documento: Article