Generation of stable integration-free pig induced pluripotent stem cells under chemically defined culture condition.
Generation of stable integration-free pig induced pluripotent stem cells under chemically defined culture condition.
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DOI：
10.1111/cpr.13487
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发表时间：
2023-11
影响因子：
8.5
通讯作者：
中科院分区：
文献类型：
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无基因组整合的猪诱导多能干细胞(IPSCs)在再生医学的临床前测试以及濒危或稀有地方猪种质资源的保护和开发中具有巨大的价值。然而，由于缺乏合适的培养液，具有实用价值的猪IPSCs的高效诱导和稳定维持仍具有挑战性。在这里，我们建立了一种高效的外源基因非依赖性IPSCs的诱导系统，该系统以前用于产生稳定的猪前原肠形成上皮干细胞(PgEpiSCs)。WNT抑制在外源基因非依赖性IPSCs的建立中起着至关重要的作用。值得注意的是，在此培养条件下，利用异体载体可以从猪体细胞中建立稳定的、无整合的猪IPSCs。IPSCs具有与pgEpiSCs相似的多能性特征和转录组特征。更重要的是，这种诱导系统可以用来从老年残疾的当地稀有猪体细胞中产生无整合的IPSCs，这些IPSCs可以经过基因编辑，用作核移植的供体细胞。我们的结果为家畜品种的遗传育种和再生医学的临床前评估提供了新的见解。本研究提出的模型概述。猪诱导的多能干细胞(IPSCs)在3I/LAF培养基下，用逆转录病毒或上体载体重编程，在没有外源基因的情况下，可以保持长期的自我更新和多能性。利用表体重编程系统，可以产生稀有的无整合的猪IPSCs，可以对其进行基因编辑，作为供体细胞进行克隆。
Genome integration‐free pig induced pluripotent stem cells (iPSCs) bring tremendous value in pre‐clinical testing of regenerative medicine, as well as conservation and exploitation of endangered or rare local pig idioplasmatic resources. However, due to a lack of appropriate culture medium, efficient induction and stable maintenance of pig iPSCs with practical value remains challenging. Here, we established an efficient induction system for exogenous gene‐independent iPSCs under chemically defined culture condition previously used for generation of stable pig pre‐gastrulation epiblast stem cells (pgEpiSCs). WNT suppression was found to play an essential role in establishment of exogenous gene‐independent iPSCs. Strikingly, stable integration‐free pig iPSCs could be established from pig somatic cells using episomal vectors in this culture condition. The iPSCs had pluripotency features and transcriptome characteristics approximating pgEpiSCs. More importantly, this induction system may be used to generate integration‐free iPSCs from elderly disabled rare local pig somatic cells and the iPSCs could be gene‐edited and used as donor cells for nuclear transfer. Our results provide novel insights into potential applications for genetic breeding of livestock species and pre‐clinical evaluation of regenerative medicine. An overview of model presented by this study. Pig induced pluripotent stem cells (iPSCs) reprogrammed using retrovirus or episomal vectors under the 3i/LAF culture medium could maintain long‐term self‐renewal and pluripotency without exogenous genes. Using episomal reprogramming system, rare pig integration‐free iPSCs could be generated, which could be gene‐edited and served as donor cells for cloning.
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影响因子：
64.8
作者：
Kobayashi T;Zhang H;Tang WWC;Irie N;Withey S;Klisch D;Sybirna A;Dietmann S;Contreras DA;Webb R;Allegrucci C;Alberio R;Surani MA
通讯作者：
Surani MA
影响因子：
64.8
作者：
Komor AC;Kim YB;Packer MS;Zuris JA;Liu DR
通讯作者：
Liu DR
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DOI：
10.1016/j.bbrc.2015.07.074
发表时间：
2015-08-28
影响因子：
3.1
作者：
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通讯作者：
Zhang, Yong
影响因子：
23.9
作者：
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通讯作者：
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影响因子：
5.9
作者：
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