Generation of Induced Pluripotent Stem Cells from Human Cord Blood Using OCT4 and SOX2

Center of Regenerative Medicine in Barcelona, Dr. Aiguader 88, Barcelona, Spain.
Cell stem cell (Impact Factor: 22.27). 10/2009; 5(4):353-7. DOI: 10.1016/j.stem.2009.09.008
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Available from: Trond Aasen, Sep 28, 2015
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    • "Behav. (2015), cells (Li et al., 2009; Zhao et al., 2010), cells present in urine (Zhou et al., 2012) and various other cell types (Aoki et al., 2010; Bar-Nur et al., 2011; Eminli et al., 2009; Giorgetti et al., 2009; Haase et al., 2009; J.B. Kim et al., 2009; Liu et al., 2010; Nakagawa et al., 2008; Sugii et al., 2011; Yu et al., 2007). Although a well-established cell type in many fields of research, due to their ease of handling and the costeffectiveness , there are disadvantages to the use of fibroblasts as a starting cell type for producing hiPSCs. "
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    ABSTRACT: Over recent years tremendous progress has been made towards understanding the molecular and cellular mechanism by which estrogens exert enhancing effects on cognition, and how they act as a neuroprotective or neurotrophic agent in disease. Currently, much of this work has been carried out in animal models with only a limited number of studies using native human tissue or cells. Recent advances in stem cell technology now make it possible to reprogram somatic cells from humans into induced pluripotent stem cells (iPSCs), which can subsequently be differentiated in neurons of specific lineages. Importantly, the reprogramming of cells allows for the generation of iPSCs that retains the genetic "makeup" of the donor. Therefore, it is possible to generate iPSC-derived neurons from patients diagnosed with specific diseases, that harbor the complex genetic background associated with the disorder. Here, we review the iPSC technology and how its currently being used to model neural development and neurological diseases. Furthermore, we explore whether this cellular system could be used to understand the role of estrogens in human neurons, and present preliminary data in support of this. We further suggest that the use of iPSC technology offers a novel system in which to not only further understand estrogens' effects in human cells, but in which to investigate the mechanism by which estrogens are beneficial in disease. Developing a greater understanding of these mechanisms in native human cells will also aid in the development of safer and more effective estrogen-based therapeutics. Copyright © 2015. Published by Elsevier Inc.
    Hormones and Behavior 07/2015; 26. DOI:10.1016/j.yhbeh.2015.06.014 · 4.63 Impact Factor
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    • "After choosing the target donor somatic cell type, it is necessary to select a cocktail of reprograming factors that usually are the four above mentioned. In few situations less than four factors are needed, such as in the case of cord blood CD133+ cells and keratinocytes (117). Through the reprograming process, the chromatin remodeling plays an essential role in the procurement of pluripotency. "
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    ABSTRACT: Natural killer (NK) cells play an essential role in the fight against tumor development. Over the last years, the progress made in the NK-cell biology field and in deciphering how NK-cell function is regulated, is driving efforts to utilize NK-cell-based immunotherapy as a promising approach for the treatment of malignant diseases. Therapies involving NK cells may be accomplished by activating and expanding endogenous NK cells by means of cytokine treatment or by transferring exogenous cells by adoptive cell therapy and/or by hematopoietic stem cell transplantation. NK cells that are suitable for adoptive cell therapy can be derived from different sources, including ex vivo expansion of autologous NK cells, unstimulated or expanded allogeneic NK cells from peripheral blood, derived from CD34+ hematopoietic progenitors from peripheral blood and umbilical cord blood, and NK-cell lines. Besides, genetically modified NK cells expressing chimeric antigen receptors or cytokines genes may also have a relevant future as therapeutic tools. Recently, it has been described the derivation of large numbers of functional and mature NK cells from pluripotent stem cells, both embryonic stem cells and induced pluripotent stem cells, which adds another tool to the expanding NK-cell-based cancer immunotherapy arsenal.
    Frontiers in Immunology 09/2014; 5:439. DOI:10.3389/fimmu.2014.00439
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    • "Thus, a faster protocol such as conversion of human patient-specific pluripotent stem cells and fibroblasts directly to retinal photoreceptors by forced expression of photoreceptor-expressed transcription factors might be more suitable to the use of these nanoparticles. Cord blood cells and keratinocytes have been shown to be more amenable for reprogramming, in terms of speed and efficiency, than skin fibroblasts.39,40 Cord blood cells have a high population of progenitor stem cells, and the keratinocyte is an epithelial cell type similar to the embryonic stem cell, whereas dermal fibroblasts (especially adult) are differentiated mesenchymal cells that are close to senescence,41,42 can possess somatic mutations,43,44 and have to undergo mesenchymal to epithelial transition during reprogramming. "
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    ABSTRACT: Gene delivery can potentially be used as a therapeutic for treating genetic diseases, including neurodegenerative diseases, as well as an enabling technology for regenerative medicine. A central challenge in many gene delivery applications is having a safe and effective delivery method. We evaluated the use of a biodegradable poly(beta-amino ester) nanoparticle-based nonviral protocol and compared this with an electroporation-based approach to deliver episomal plasmids encoding reprogramming factors for generation of human induced pluripotent stem cells (hiPSCs) from human fibroblasts. A polymer library was screened to identify the polymers most promising for gene delivery to human fibroblasts. Feeder-independent culturing protocols were developed for nanoparticle-based and electroporation-based reprogramming. The cells reprogrammed by both polymeric nanoparticle-based and electroporation-based nonviral methods were characterized by analysis of pluripotency markers and karyotypic stability. The hiPSC-like cells were further differentiated toward the neural lineage to test their potential for neurodegenerative retinal disease modeling. 1-(3-aminopropyl)-4-methylpiperazine end-terminated poly(1,4-butanediol diacry-late-co-4-amino-1-butanol) polymer (B4S4E7) self-assembled with plasmid DNA to form nanoparticles that were more effective than leading commercially available reagents, including Lipofectamine® 2000, FuGENE® HD, and 25 kDa branched polyethylenimine, for nonviral gene transfer. B4S4E7 nanoparticles showed effective gene delivery to IMR-90 human primary fibroblasts and to dermal fibroblasts derived from a patient with retinitis pigmentosa, and enabled coexpression of exogenously delivered genes, as is needed for reprogramming. The karyotypically normal hiPSC-like cells generated by conventional electroporation, but not by poly(beta-amino ester) reprogramming, could be differentiated toward the neuronal lineage, specifically pseudostratified optic cups. This study shows that certain nonviral reprogramming methods may not necessarily be safer than viral approaches and that maximizing exogenous gene expression of reprogramming factors is not sufficient to ensure successful reprogramming.
    International Journal of Nanomedicine 12/2013; 8:4641-4658. DOI:10.2147/IJN.S53830 · 4.38 Impact Factor
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