Article

Generation of Induced Pluripotent Stem Cells with CytoTune, a Non-Integrating Sendai Virus

Primary and Stem Cell Systems, Life Technologies, Carlsbad, CA, USA.
Methods in molecular biology (Clifton, N.J.) (Impact Factor: 1.29). 04/2013; 997:45-56. DOI: 10.1007/978-1-62703-348-0_5
Source: PubMed

ABSTRACT

One of the major obstacles in generating induced pluripotent stem cells for research or downstream applications is the potential modifications of cellular genome as a result of using integrating viruses during reprogramming. Another major disadvantage of reprogramming cells with integrating vectors is that silencing and activation of transgenes are unpredictable, which may affect terminal differentiation potential and increase the risk of using iPSC-derived cells. Here we describe a protocol for the generation of induced pluripotent stem cells using a non-integrating RNA virus, Sendai virus, to efficiently generate transgene-free iPSCs starting with different cell types as well as in feeder-free conditions.

Download full-text

Full-text

Available from: Mohan Vemuri, Jul 14, 2015
  • Source
    • "To eliminate the possible effects of insertion and mutagenesis, a lot of modified reprogramming methods have been reported [72], such as non-integration adenoviral102103104105 and sendai viral106107108 vector, episomal vector109110111, minicircle [112], plasmid [113], PiggyBac transposon [114], messenger RNAs [115] and miRNAs [116] and excisable expression systems [117, 118]. Compared with viral methods, nucleic acid based approaches mentioned above are much safer, but these methods still cannot get rid of all the possible risk of tumorigenesis. "
    [Show abstract] [Hide abstract]
    ABSTRACT: The plasma membrane remains a major barrier for intracellular drug delivery, to overcome this issue, a variety of approaches have been developed and used to deliver therapeutic cargos. Among these approaches, cell penetrating peptide (CPP) is promising and affords widely used vector for efficient intracellular delivery of cargos. Moreover, the latter findings including iPS reprogramming and direct transdifferentiation as well as gene editing have gradually become hot research topic; because their application in tissue engineering and disease modeling have great potential to advance innovation in precision medicine. Since the beginning, research on these approaches is mainly based on virus transduction system, while, under the consideration for obviating the risk of mutagenic insertion and enables more accurate controlling, CPP-based efficient virus-free delivery strategy has been used recently. In this review, we summarize the existing CPP-based delivery system, emerging landscape of CPP application in stem cell manipulation and reprogramming, along with CPP contributions to gene editing techniques.
    Full-text · Article · Feb 2016 · Journal of Controlled Release
    • "iPSCs have the capacity to differentiate into patient specific cell types, therefore reducing the likelihood of immunological rejection. Since the first report by Nobel Prize laureate Yamanaka that iPSCs could be generated from fibroblasts through the introduction of specific transcription factors using retroviral systems (Takahashi et al., 2007; Takahashi and Yamanaka, 2006), more effective, non-integrative techniques involving the use of Sendai virus have been described (Lieu et al., 2013). Thus, it is currently possible to generate iPSCs from a person's somatic cells and differentiate them into the required functional cell type (Wilson and Wu, 2015): notably, a first human clinical trial is using iPSC-derived retinal pigment epithelium cells to evaluate their efficacy in treating macular degeneration (Reardon and Cyranoski, 2014). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Induced pluripotent stem cells (iPSCs) are a promising tool for regenerative medicine in chronic conditions associated with muscle atrophy since iPSCs are easier to obtain, pose less ethical limitations and can better capture human genetic diversity compared with human embryonic stem cells. We highlight the potentiality of iPSCs for treating muscle-affecting conditions for which no effective cure is yet available, notably aging sarcopenia and inherited neurometabolic conditions. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
    No preview · Article · Jul 2015 · Journal of Cellular Physiology
  • Source
    • "Induced PSC were generated from freshly collected, peripheral blood-derived PBMC (Chou et al., 2011). Cells were transduced with the integration-free CytoTune-iPS Sendai Reprogramming Kit, which utilizes Sendai virus particles of the four Yamanaka factors (Lieu et al., 2013; Takahashi et al., 2007). Transduced cells were plated with MEF feeder cells, and fed iPSC medium with freshly added bFGF (Invitrogen) until small colonies were formed in about 2 weeks. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Peripheral blood mononuclear cells (PBMC) were donated by a patient with clinically diagnosed frontotemporal dementia (FTD). Induced pluripotent stem cells (iPSCs) were developed using integration-free CytoTune-iPS Sendai Reprogramming factors which include Sendai virus particles of the four Yamanaka factors Oct, Sox2, Klf4, and c-Myc. Published by Elsevier B.V.
    Full-text · Article · Jul 2015 · Stem Cell Research
Show more