Exosome-mediated direct conversion of human fibroblasts into neural progenitor-like cells
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ABSTRACT:
PROVIDER: PRJNA384018 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR5470270.fastq.gz | Fastqsanger.gz | |||
| SRR5470271.fastq.gz | Fastqsanger.gz | |||
| SRR5470272.fastq.gz | Fastqsanger.gz | |||
| SRR5470292_1.fastq.gz | Fastqsanger.gz | |||
| SRR5470292_2.fastq.gz | Fastqsanger.gz |
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Project description:Exosome, a kind of extracellular vesicles, has been introduced as a vehicle for horizontal transferring of genetic material. We induced autologous exosome containing a cocktail of reprogramming factors (‘reprosome’) to convert fibroblasts into neural progenitor cells (NPC). The fibroblasts were sonicated and subsequently cultured in NSC media for 1 day to induce release of reprosomes composed of reprogramming factors associated to chromatin remodeling and neural lineage-specific factors. After treated with reprosomes, fibroblasts were converted into NPCs (rNPC) with great efficiency via activation of chromatin remodeling, as fast as it required only five days for formation of 1,500 spheroids showing NPC-like phenotype. The rNPCs maintained self-renewal and proliferative properties for several weeks and successfully differentiated into neuron, astrocyte, and oligodendrocyte, in vitro and in vivo. Reprosome-mediated cellular reprogramming is simple, safe, and efficient, thus providing a new paradigm for clinical application that requires autologous stem cells.
2018-04-24 | GSE98113 | GEO
Project description:Exosome, a kind of extracellular vesicles, has been introduced as a vehicle for horizontal transferring of genetic material. We induced autologous exosome containing a cocktail of reprogramming factors (‘reprosome’) to convert fibroblasts into neural progenitor cells (NPC). The fibroblasts were sonicated and subsequently cultured in NSC media for 1 day to induce release of reprosomes composed of reprogramming factors associated to chromatin remodeling and neural lineage-specific factors. After treated with reprosomes, fibroblasts were converted into NPCs (rNPC) with great efficiency via activation of chromatin remodeling, as fast as it required only five days for formation of 1,500 spheroids showing NPC-like phenotype. The rNPCs maintained self-renewal and proliferative properties for several weeks and successfully differentiated into neuron, astrocyte, and oligodendrocyte, in vitro and in vivo. Reprosome-mediated cellular reprogramming is simple, safe, and efficient, thus providing a new paradigm for clinical application that requires autologous stem cells.
2018-04-24 | GSE98111 | GEO
Project description:Exosome, a kind of extracellular vesicles, has been introduced as a vehicle for horizontal transferring of genetic material. We induced autologous exosome containing a cocktail of reprogramming factors (‘reprosome’) to convert fibroblasts into neural progenitor cells (NPC). The fibroblasts were sonicated and subsequently cultured in NSC media for 1 day to induce release of reprosomes composed of reprogramming factors associated to chromatin remodeling and neural lineage-specific factors. After treated with reprosomes, fibroblasts were converted into NPCs (rNPC) with great efficiency via activation of chromatin remodeling, as fast as it required only five days for formation of 1,500 spheroids showing NPC-like phenotype. The rNPCs maintained self-renewal and proliferative properties for several weeks and successfully differentiated into neuron, astrocyte, and oligodendrocyte, in vitro and in vivo. Reprosome-mediated cellular reprogramming is simple, safe, and efficient, thus providing a new paradigm for clinical application that requires autologous stem cells.
2018-04-24 | GSE98110 | GEO
Project description:Exosome, a kind of extracellular vesicles, has been introduced as a vehicle for horizontal transferring of genetic material. We induced autologous exosome containing a cocktail of reprogramming factors (‘reprosome’) to convert fibroblasts into neural progenitor cells (NPC). The fibroblasts were sonicated and subsequently cultured in NSC media for 1 day to induce release of reprosomes composed of reprogramming factors associated to chromatin remodeling and neural lineage-specific factors. After treated with reprosomes, fibroblasts were converted into NPCs (rNPC) with great efficiency via activation of chromatin remodeling, as fast as it required only five days for formation of 1,500 spheroids showing NPC-like phenotype. The rNPCs maintained self-renewal and proliferative properties for several weeks and successfully differentiated into neuron, astrocyte, and oligodendrocyte, in vitro and in vivo. Reprosome-mediated cellular reprogramming is simple, safe, and efficient, thus providing a new paradigm for clinical application that requires autologous stem cells.
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Project description:Many studies have already shown the reprogramming of somatic cells into other cell types such as neural stem cells, blood progenitor cells, and hepatocytes by inducing combinations of transcription factors. One of the recent development in cellular reprogramming is the direct reprogramming, that can change cell fate towards different lineages. This strategy provides an alternative to the use of pluripotent stem cells ruling out the concerns of tumorigenicity caused by undifferentiated cell populations. Here, we generated induced oligodendrocyte progenitor cells (iOPCs) from mouse fibroblasts by direct reprogramming. The generated iOPCs are homogenous, self-renewing, and multipotent. Once differentiated, the somatic stem cells exhibit morphological and molecular characteristics of oligodendrocyte progenitor cells (OPCs). Thus, we demonstrated that terminally differentiated somatic cells can be converted into functional iOPCs by induction of transcription factors offering a new strategies to cure myelin disorders. To identify the global gene expression profiles of iOPCs, we analyzed total 6 samples.
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Project description:Many studies have already shown the reprogramming of somatic cells into other cell types such as neural stem cells, blood progenitor cells, and hepatocytes by inducing combinations of transcription factors. One of the recent development in cellular reprogramming is the direct reprogramming, that can change cell fate towards different lineages. This strategy provides an alternative to the use of pluripotent stem cells ruling out the concerns of tumorigenicity caused by undifferentiated cell populations. Here, we generated induced oligodendrocyte progenitor cells (iOPCs) from mouse fibroblasts by direct reprogramming. The generated iOPCs are homogenous, self-renewing, and multipotent. Once differentiated, the somatic stem cells exhibit morphological and molecular characteristics of oligodendrocyte progenitor cells (OPCs). Thus, we demonstrated that terminally differentiated somatic cells can be converted into functional iOPCs by induction of transcription factors offering a new strategies to cure myelin disorders.
2015-11-02 | GSE52035 | GEO
Project description:Direct lineage conversion holds great promise in the regenerative medicine field for restoring damaged tissues using functionally engineered counterparts. However, current methods of direct lineage conversion, even those employing virus-mediated transgenic expression of tumorigenic factors, are extremely inefficient (~25%). Thus, advanced methodologies capable of revolutionizing efficiency and addressing safety issues are key to clinical translation of these technologies. Here, we propose an exosome-guided, non-viral, direct-lineage conversion strategy to enhance transdifferentiation of fibroblasts to induced cardiomyocyte-like cells (iCMs). Exosomes produced during the cardiac differentiation process of embryonic stem cells (ESCs) are able to achieve extremely high reprogramming efficiency (>60%) by generating functional iCMs from mouse embryonic fibroblasts via a cardiac precursor-like stage rather than a pluripotent state. The resulting iCMs possess typical cardiac Ca2+ transients and electrophysiological features, and exhibit global gene expression profiles similar to those of cardiomyocytes. The optimized reprogramming conditions produce beating iCM clusters ~3-fold more efficiently than conventional methods. This is the first demonstration of the use of exosomes derived from ESCs undergoing cardiac differentiation as biomimetic tools to induce direct cardiac reprogramming with greatly improved efficiency, establishing a general, more readily accessible platform for broadly generating a variety of specialized somatic cells through direct lineage conversion.
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Project description:Direct lineage conversion holds great promise in the regenerative medicine field for restoring damaged tissues using functionally engineered counterparts. However, current methods of direct lineage conversion, even those employing virus-mediated transgenic expression of tumorigenic factors, are extremely inefficient (~25%). Thus, advanced methodologies capable of revolutionizing efficiency and addressing safety issues are key to clinical translation of these technologies. Here, we propose an exosome-guided, non-viral, direct-lineage conversion strategy to enhance transdifferentiation of fibroblasts to induced cardiomyocyte-like cells (iCMs). Exosomes produced during the cardiac differentiation process of embryonic stem cells (ESCs) are able to achieve extremely high reprogramming efficiency (>60%) by generating functional iCMs from mouse embryonic fibroblasts via a cardiac precursor-like stage rather than a pluripotent state. The resulting iCMs possess typical cardiac Ca2+ transients and electrophysiological features, and exhibit global gene expression profiles similar to those of cardiomyocytes. The optimized reprogramming conditions produce beating iCM clusters ~3-fold more efficiently than conventional methods. This is the first demonstration of the use of exosomes derived from ESCs undergoing cardiac differentiation as biomimetic tools to induce direct cardiac reprogramming with greatly improved efficiency, establishing a general, more readily accessible platform for broadly generating a variety of specialized somatic cells through direct lineage conversion.
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Project description:Time course micro array experiment to identify transcriptional changes in response to exposure of hFLs to different combinations of small molecules during direct neuronal reprogramming hFL1 cells were transduced with conversion factors Ascl1, Brn2a and Myt1L and five days after transduction, transgene expression was activated by doxycycline. On day 3 of transgene expression, MEF medium was replaced by N2B27 medium plus small molecules, depending on the condition. Ascl1 expression was linked to GFP so that all GFP pos cells were FACS isolated at different times of exposure to small molecules (or N2B27 medium in transgene only groups). Post FACS, cells were lysed and total RNA was extracted and used as input for micro array experiments.
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