Direct reprogramming of mouse fibroblasts to neural progenitors. Proc Natl Acad Sci USA

Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037, USA.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 05/2011; 108(19):7838-43. DOI: 10.1073/pnas.1103113108
Source: PubMed


The simple yet powerful technique of induced pluripotency may eventually supply a wide range of differentiated cells for cell therapy and drug development. However, making the appropriate cells via induced pluripotent stem cells (iPSCs) requires reprogramming of somatic cells and subsequent redifferentiation. Given how arduous and lengthy this process can be, we sought to determine whether it might be possible to convert somatic cells into lineage-specific stem/progenitor cells of another germ layer in one step, bypassing the intermediate pluripotent stage. Here we show that transient induction of the four reprogramming factors (Oct4, Sox2, Klf4, and c-Myc) can efficiently transdifferentiate fibroblasts into functional neural stem/progenitor cells (NPCs) with appropriate signaling inputs. Compared with induced neurons (or iN cells, which are directly converted from fibroblasts), transdifferentiated NPCs have the distinct advantage of being expandable in vitro and retaining the ability to give rise to multiple neuronal subtypes and glial cells. Our results provide a unique paradigm for iPSC-factor-based reprogramming by demonstrating that it can be readily modified to serve as a general platform for transdifferentiation.

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Available from: Janghwan Kim
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    • "stem cells or their progeny, as a result of possible iPSC contamination or incomplete reprogramming. This suggests that there are several limitations to the use of iPSCs in neural cell-based therapy or in neural disease modeling. In general, reprogramming factors such as the Yamanaka factors are used to drive somatic cells into a pluripotent state. Kim et al. (2011) showed that mouse fibroblasts are converted to neural stem cells by bypassing the iPSC stage with help from the same factors. Therefore, our study might also support the hypothesis that neural stem cells can be derived during the iPSC induction process. Unexpectedly, LD-iNSC colonies appeared approximately after day 14 in this study, wh"
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    ABSTRACT: In mice, leukemia inhibitory factor (LIF)-dependent primitive neural stem cells (NSCs) have a higher neurogenic potential than bFGF-dependent definitive NSCs. Therefore, expandable primitive NSCs are required for research and for the development of therapeutic strategies for neurological diseases. There is a dearth of suitable techniques for the generation of human long-term expandable primitive NSCs. Here, we have described a method for the conversion of human fibroblasts to LIF-dependent primitive NSCs using a strategy based on techniques for the generation of induced pluripotent stem cells (iPSCs). These LIF-dependent induced NSCs (LD-iNSCs) can be expanded for >100 passages. Long-term cultured LD-iNSCs demonstrated multipotent neural differentiation potential and could generate motor neurons and dopaminergic neurons, as well as astrocytes and oligodendrocytes, indicating a high level of plasticity. Furthermore, LD-iNSCs easily reverted to human iPSCs, indicating that LD-iNSCs are in an intermediate iPSC state. This method may facilitate the generation of patient-specific human neurons for studies and treatment of neurodegenerative diseases.
    Preview · Article · Oct 2015 · Biology Open
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    • "Direct conversion of somatic cells to lineage-committed stem/ progenitor cells, such as neural stem/progenitor cells (NSCs/NPCs), would allow production of sufficient cells for downstream research or application and overcome the potential risk for tumor formation by iPSCs. A more general lineage conversion approach of somatic cells has been developed using transient expression of iPSCinducing transcription factors when cultured in the presence of lineage-specifying extracellular cues [1] [2], indicating that cell fate changes of fibroblasts overexpressing Yamanaka factors are closely associated with the culture condition. In contrast, fibroblasts overexpressing neural transcription factors lacked the plasticity to generate different tissue cell fates [3e5]. "
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    ABSTRACT: Non-human primates provide optimal models for the development of stem cell therapies. Although somatic cells have been converted into neural stem/progenitor cells, it is unclear whether telencephalic neuroepithelial stem cells (NESCs) with stable properties can be generated from fibroblasts in primate. Here we report that a combination of transcription factors (Oct4, Sox2, Klf4) with a new culture medium induces rhesus monkey fibroblasts into NESCs, which can develop into miniature neural tube (NT)-like structures at a cell level. Furthermore, single induced NESCs (iNESCs) can generate later-stage 3D-NTs after grown on matrigel in suspension culture. iNESCs express NT cell markers, have a unique gene expression pattern biasing towards telencephalic patterning, and give rise to cortical neurons. Via transplantation, single iNESCs can extensively survive, regenerate myelinated neuron axons and synapse structures in adult monkey striatum and cortex, and differentiate into cortical neurons. Successful transplantation is closely associated with graft regions and grafted cell identities. The ability to generate defined and transplantable iNESCs from primate fibroblasts under a defined condition with predictable fate choices will facilitate disease modeling and cell therapy.
    Full-text · Article · Oct 2015 · Biomaterials
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    • "More recently, somatic cells were redirected into another differentiated cell lineage, without passing an intermediate pluripotent stage, by transducing a set of transcription factors that play crucial regulatory roles in the differentiation of the destination cell; direct conversion , or direct reprogramming, of murine fibroblasts into cardiomyocytes (Ieda et al., 2010; Inagawa et al., 2012; Inagawa and Ieda, 2013), neurons (Kim et al., 2011, 2012; Caiazzo et al., 2011; Han et al., 2012), chondrocytes (Hiramatsu et al., 2011), and hepatocytes (Sekiya and Suzuki, 2011; Huang et al., 2011), as well as of human fibroblasts into cardiomyocytes (Wada et al., 2013; Nam et al., 2013), neurons (Pang et al., 2011; Caiazzo et al., 2011; Kim et al., 2012), and hematopoietic cells (Szabo et al., 2010) have been reported. Although the efficiencies of direct conversion were generally low (0.005%–30% of fibroblasts were successfully converted into the cells of interest; Thier et al., 2012; Sekiya and Suzuki, 2011; Pang et al., 2011; Nam et al., 2013; Kim et al., 2011, 2012; Inagawa et al., 2012; Inagawa and Ieda, 2013; Ieda et al., 2010; Huang et al., 2011; Han et al., 2012; Caiazzo et al., 2011), these technologies may be quite valuable for generation of the desired cells that could be used in basic research as well as in regenerative therapy for various human disorders. We have recently succeeded in directly reprogramming human fibroblasts into osteoblasts that massively produced bone matrix and contributed to bone regeneration with an efficiency as high as 80% (Yamamoto et al., 2015). "
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    ABSTRACT: Brown adipocytes (BAs) play important roles in body temperature regulation, energy balance, and carbohydrate and lipid metabolism. Activities of BAs are remarkably diminished in obese and diabetic patients, providing possibilities of transplanting functional BAs resulting in therapeutic benefit. Here, we show generation of functional BAs by cellular reprogramming procedures. Transduction of the PRDM16 gene into iPSC-derived embryoid bodies induced BA phenotypes (iBAs). Moreover, normal human fibroblasts were directly converted into BAs (dBAs) by C/EBP-β and C-MYC gene transduction. Approximately 90% of the fibroblasts were successfully converted within 12 days. The dBAs were highly active in mitochondrial biogenesis and oxidative metabolism. Mouse dBAs were induced by Prdm16, C/ebp-β, and L-myc genes, and after transplantation, they significantly reduced diet-induced obesity and insulin resistance in an UCP1-dependent manner. Thus, highly functional BAs can be generated by cellular reprogramming, suggesting a promising tailor-made cell therapy against metabolic disorders including type 2 diabetes mellitus.
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