Direct reprogramming of fibroblasts into a cardiovascular tissue
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ABSTRACT:
PROVIDER: PRJNA744237 | ENA |
REPOSITORIES: ENA
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Project description:Purpose: To explore detailed transcriptome changes during the transition from fibroblasts toward cardiac cells Methods: We performed single-cell RNA-seq on MTTFs (11,580 cells) and MTTFs treated with MAB for 6 days (12,261 cells). In MTTFs, the mean reads and mediam genes per cell were 53,107 and 5,096, respectively; In reprogrammed cells, the mean reads and medium genes per cell were 100,515 and 6,164, respectively. Result & Conclusion: Single-cell RNA-seq studies demonstrate a simultaneous reprogramming of mouse tail-tip fibroblasts (MTTFs) by MAB into CMs, Ecs and SMCs.
2021-07-08 | GSE179589 | GEO
Project description:Direct reprogramming of mouse fibroblasts into functional skeletal muscle progenitors
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Project description:Mouse tail-tip fibroblasts could be reprogrammed toward cardiomimetic tissue by combined treatments of miRNA, ascorbic acid and BMP4
2020-03-13 | GSE96617 | GEO
Project description:Ultra-Efficient Exosome-Guided Direct Reprogramming of Fibroblasts into Functional Cardiomyocytes
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Project description:We developed a strategy to capture cardiovascular progenitor cells during the cardiac reprogramming from mouse fibroblasts and expand these CPCs for a long term in a chemically defined condition.
2016-01-29 | GSE77375 | GEO
Project description:We report differential mRNA expression levels in primary mouse embryonic fibroblasts, primary neurons, and induced neuronal cells generated via direct reprogramming method. The induced neuronal cells were cultured on laminin or decellularized brain ECM.
2018-07-01 | GSE114809 | 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.
2022-03-02 | GSE166250 | 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.
2022-03-02 | GSE166249 | GEO
Project description:Recent advances in the stem cell biology have revealed that cell type-specific transcription factors could reset the somatic memory and induce direct reprogramming into specific cellular identities. The induction of pluripotency in terminally differentiated cells has been a major achievement in the field of direct reprogramming. Recent studies have shown that fibroblasts could be directly converted into specific cell types, such as neurons, cardiomyocytes, blood progenitor cells, and epiblast stem cells, without first passing through an induced pluripotent stem cell state3-7. However, direct reprogramming of differentiated cells into somatic stem cell types has not been described yet4. Here we show that a combination of neural-specific transcription factors (Sox2, Klf4, c-Myc, Brn4/Pou3f4 or Sox2, Klf4, c-Myc, Brn4, E47/Tcf3) can induce a neural stem cell (NSC) fate on the fibroblasts. Induced neural stem cells (iNSCs) showed morphology, gene expression, epigenetic features, differentiation potential, and functionality similar to wild-type NSCs. Therefore, our data suggest that cell type-specific defined factors can induce specific stem cell identities on somatic cells. Fibroblasts (5 x 104 cells) were infected with retroviruses for two days, and cells were maintained in NSC media: DMEM/F-12 supplemented with N2 or B27 supplements (Gibco-BRL), 10 ng/ml EGF, 10 ng/ml bFGF (both from Invitrogen), 50 ug/ml BSA (Fraction V Gibco-BRL), and 1x penicillin/streptomycin/glutamine (Gibco-BRL). In order to establish stable iNSC lines, were either manually picked mature iNSC clumps or passaged and seeded the whole dishes of cells onto either gelatin- or laminin-coated dishes. RNA samples to be analysed on microarrays were prepared using QIAGEN RNeasy columns with on-column DNA digestion. 500 ng of total RNA per sample was used as input RNA into a linear amplification protocol (Ambion) involving synthesis of T7-linked double-stranded cDNA and 12 h of in-vitro transcription incorporating biotin-labelled nucleotides. Purified and labelled cRNA was hybridised onto MouseRef-8 v2 expression BeadChips (Illumina) for 18 h according to the manufacturer's instructions. After washing, as recommended, chips were stained with streptavidin-Cy3 (GE Healthcare) and scanned using iScan reader (Illumina) and accompanying software. Samples were hybridised as biological replicates. 9 samples were analyzed. Fibroblast: CF1 Mouse embryonic fibroblasts, 1 biological rep Control NSC: OG2-Rosa Neural Stem Cells, 2 biological rep 4F iNSC (late): 4 factors induced Neural Stem Cells (late passage), 2 biological rep 4F iNSC (early): 4 factors induced Neural Stem Cells (early passage), 2 biological rep 5F iNSC: 5 factors induced Neural Stem Cells, 2 biological rep
2012-05-02 | E-GEOD-30500 | biostudies-arrayexpress
Project description:Direct reprogramming of fibroblasts into cardiomyocyte-like cells (iCM) holds great potential for heart regeneration and disease modeling and may lead to future therapeutic applications in human patients with heart disease. Currently, the application of this technology is limited by our lack of understanding of the molecular mechanisms which drive direct iCM reprogramming. Using a quantitative mass spectrometry-based proteomic approach we have identified the temporal global changes in protein abundance that occur during the initial phases of iCM reprogramming. Collectively, our results show systematic and temporally distinct alterations in the levels of specific functional classes of proteins during the initiating steps of reprogramming including extracellular matrix proteins, translation factors, and chromatin-binding proteins.
2018-02-19 | PXD006152 | Pride