Generation of Rat and Human Induced Pluripotent Stem Cells by Combining Genetic Reprogramming and Chemical Inhibitors

Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Cell stem cell (Impact Factor: 22.27). 04/2009; 4(1):16-9. DOI: 10.1016/j.stem.2008.11.014
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Available from: Saiyong Zhu, Jun 13, 2014
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    • "Human or non-human primate PSCs with some features of naïve pluripotency have been derived by the conversion of conventional hESCs or induced PSCs (iPSCs) in vitro, through reprogramming of somatic cells to a pluripotent state or by direct derivation from the embryo (Tables 2 and 3) (Buecker et al., 2010; Chan et al., 2013; Fang et al., 2014; Gafni et al., 2013; Hanna et al., 2010; Li et al., 2009; Takashima et al., 2014; Theunissen et al., 2014; Valamehr et al., 2014; Ware et al., 2014, Wang et al., 2014). Early studies used exogenous transgenes to trigger the conversion from primed to naïve pluripotency and, in some cases, to stabilise and maintain the naïve state. "
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    ABSTRACT: In the mouse, naïve pluripotent stem cells (PSCs) are thought to represent the cell culture equivalent of the late epiblast in the pre-implantation embryo, with which they share a unique defining set of features. Recent studies have focused on the identification and propagation of a similar cell state in human. Although the capture of an exact human equivalent of the mouse naïve PSC remains an elusive goal, comparative studies spurred on by this quest are lighting the path to a deeper understanding of pluripotent state regulation in early mammalian development.
    Development 09/2015; 142:30910-3099. DOI:10.1242/dev.116061 · 6.46 Impact Factor
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    • "Chenxia Hu, Lanjuan Li molecules that affect methylation or acetylation, mimic the Wnt-signaling pathway, or modulate the TGF-b pathway (Li et al., 2009). These methods raised few ethical concerns because of their derivation from somatic cells and, thus, are powerful tools for studying basic developmental biology. "
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    ABSTRACT: Various liver diseases result in terminal hepatic failure, and liver transplantation, cell transplantation and artificial liver support systems are emerging as effective therapies for severe hepatic disease. However, all of these treatments are limited by organ or cell resources, so developing a sufficient number of functional hepatocytes for liver regeneration is a priority. Liver regeneration is a complex process regulated by growth factors (GFs), cytokines, transcription factors (TFs), hormones, oxidative stress products, metabolic networks, and microRNA. It is well-known that the function of isolated primary hepatocytes is hard to maintain; when cultured in vitro, these cells readily undergo dedifferentiation, causing them to lose hepatocyte function. For this reason, most studies focus on inducing stem cells, such as embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs), hepatic progenitor cells (HPCs), and mesenchymal stem cells (MSCs), to differentiate into hepatocyte-like cells (HLCs) in vitro. In this review, we mainly focus on the nature of the liver regeneration process and discuss how to maintain and enhance in vitro hepatic function of isolated primary hepatocytes or stem cell-derived HLCs for liver regeneration. In this way, hepatocytes or HLCs may be applied for clinical use for the treatment of terminal liver diseases and may prolong the survival time of patients in the near future.
    Protein & Cell 06/2015; 6(8). DOI:10.1007/s13238-015-0180-2 · 3.25 Impact Factor
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    • "Engineered endonucleases, including zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system, are invaluable tools for the rapid generation of GM animals including rats5678. Importantly, these new technologies provide genome-editing approaches for a wide variety of organisms that were previously inaccessible without embryonic stem (ES) cells91011 and induced pluripotent stem (iPS) cells1213. GM animals are usually produced by microinjecting engineered endonucleases into pronuclear-stage embryos56. Although this method is now the gold standard, it requires sophisticated manual skills to prevent cell damage. "
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    ABSTRACT: Engineered endonucleases, such as zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system, provide a powerful approach for genome editing in animals. However, the microinjection of endonucleases into embryos requires a high skill level, is time consuming, and may cause damage to embryos. Here, we demonstrate that the electroporation of endonuclease mRNAs into intact embryos can induce editing at targeted loci and efficiently produce knockout rats. It is noteworthy that the electroporation of ZFNs resulted in an embryonic survival rate (91%) and a genome-editing rate (73%) that were more than 2-fold higher than the corresponding rates from conventional microinjection. Electroporation technology provides a simple and effective method to produce knockout animals.
    Scientific Reports 10/2014; 4:6382. DOI:10.1038/srep06382 · 5.58 Impact Factor
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