Homo sapiens
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ABSTRACT:
PROVIDER: PRJNA188388 | ENA |
REPOSITORIES: ENA
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Project description:BACKGROUND: Cell-based regeneration therapies hold great promise and potential for new area in clinical medicine, although some obstacles still remain to be overcome for a wide range of clinical applications. One of the major impediments in this field is difficulties in large-scale production of cells of interest with reproducibility. Current protocol of cell therapy requires a long-time laborious manual process. To solve this problem, we focused on the robotics of automated and high throughput cell culture system. To date, an automated robotic cultivation of stem or progenitor cells in clinical trials has not been reported. METHODS: The system, AutoCulture®, used in this study can automatically replace the medium, centrifuge cells, split the cells, and take a photograph for their morphology. We examined the feasibility to use it in the clinical field by comparing the growth rate and the characteristics of cardiac stem cells (CSCs) cultivated by AutoCulture and the manual handling culture, which protocol is the exact same as current performing clinical trial in our institute. RESULTS: We demonstrated similar characteristics in both culture methods, in terms of growth rates, gene expression profiles, cell surface profiles by FACS, carbon hydrate structures on the cell surface, and genomic DNA stability. IMPLICATIONS: The results of this study showed that AutoCulture is feasible to cultivate human cells for regenerative medicine. An automated cell-processing machine for cell therapy will play more important role, as it will be widespread from multi-center trials to off-the-shelf cell products. The cells were cultured by manual handling or by the automatic cell culture apparatus. Total RNA was extracted from these cells at day 7 by the RNeasy Plus Mini Kit (QIAGEN). Gene expression analysis was performed using the Agilent Whole Human Genome Microarray chips G4112F (Agilent). Raw data were normalized and analyzed by GeneSpring GX11 software.
2013-03-31 | E-GEOD-44032 | biostudies-arrayexpress
Project description:BACKGROUND: Cell-based regeneration therapies hold great promise and potential for new area in clinical medicine, although some obstacles still remain to be overcome for a wide range of clinical applications. One of the major impediments in this field is difficulties in large-scale production of cells of interest with reproducibility. Current protocol of cell therapy requires a long-time laborious manual process. To solve this problem, we focused on the robotics of automated and high throughput cell culture system. To date, an automated robotic cultivation of stem or progenitor cells in clinical trials has not been reported. METHODS: The system, AutoCulture®, used in this study can automatically replace the medium, centrifuge cells, split the cells, and take a photograph for their morphology. We examined the feasibility to use it in the clinical field by comparing the growth rate and the characteristics of cardiac stem cells (CSCs) cultivated by AutoCulture and the manual handling culture, which protocol is the exact same as current performing clinical trial in our institute. RESULTS: We demonstrated similar characteristics in both culture methods, in terms of growth rates, gene expression profiles, cell surface profiles by FACS, carbon hydrate structures on the cell surface, and genomic DNA stability. IMPLICATIONS: The results of this study showed that AutoCulture is feasible to cultivate human cells for regenerative medicine. An automated cell-processing machine for cell therapy will play more important role, as it will be widespread from multi-center trials to off-the-shelf cell products.
2013-03-31 | GSE44032 | GEO
Project description:Efficient generation of functional cardiomyocytes from human induced pluripotent stem cells (hiPSC-CMs) is critical for their use in regenerative medicine and other applications. In this study, we evaluated the effect of space microgravity (µg) on the differentiation of hiPSC-derived cardiac progenitors compared with parallel 1g condition on the International Space Station. Cryopreserved 3D cardiac progenitors derived from hiPSCs were cultured for 3 weeks. Compared with 1g culture, the µg culture had larger sphere sizes, increased expression of proliferation markers, higher counts of nuclei, and higher cell viability. Highly enriched cardiomyocytes generated in µg had appropriate gene expression and cardiac structure as well as improved function including contractility and Ca2+ handling. RNA-seq analysis of 3-day cultures revealed that short-term exposure of cardiac progenitor spheres to space microgravity upregulated genes involved in cell proliferation, cardiac differentiation, and contraction. These results indicate that space microgravity increased survival and proliferation of hiPSC-CMs and improved their structures and functions.
2022-09-08 | GSE188793 | GEO
Project description:The mammalian heart undergoes maturation during postnatal life to meet the increased functional requirements of the adult. However, the key drivers of this process remain poorly defined. We developed as 96-well screening platform, using human pluripotent stem cell derived cardiac organoids, to determine the molecular requirements for in vitro cardiomyocyte maturation. Here, we describe gene expression changes resulting from culturing human cardiac organoids in standard cell culture conditions and under optimized maturation conditions. We assessed our maturation conditions by comparing transcriptional changes of human cardiac organoids to RNA isolated from human heart. Interesting, analysis of these data revealed that a switch to fatty acid oxidative metabolism is a key governor of cardiomyocyte maturation and mature cardiac organoids were refractory to mitogenic stimuli.
2017-08-30 | GSE93841 | GEO
Project description:Background: We had shown that cardiomyocytes (CMs) were more efficiently differentiated from human induced pluripotent stem cells (hiPSCs) when the hiPSCs were reprogrammed from cardiac fibroblasts rather than dermal fibroblasts or blood mononuclear cells. Here, we continued to investigate the relationship between somatic-cell lineage and hiPSC-CM production by comparing the yield and functional properties of CMs differentiated from iPSCs reprogrammed from human atrial or ventricular cardiac fibroblasts (AiPSC or ViPSC, respectively). Methods: Atrial and ventricular heart tissues were obtained from the same patient, reprogrammed into AiPSCs or ViPSCs, and then differentiated into CMs (AiPSC-CMs or ViPSC-CMs, respectively) via established protocols. Results: The time-course of expression for pluripotency genes (OCT4, NANOG, and SOX2), the early mesodermal marker Brachyury, the cardiac mesodermal markers MESP1 and Gata4, and the cardiovascular progenitor-cell transcription factor NKX2.5 were broadly similar in AiPSC-CMs and ViPSC-CMs during the differentiation protocol. Flow-cytometry analyses of cardiac troponin T expression also indicated that purity of the two differentiated hiPSC-CM populations (AiPSC-CMs: 88.23±4.69%, ViPSC-CMs: 90.25±4.99%) was equivalent. While the field-potential durations were significantly longer in ViPSC-CMs than in AiPSC-CMs, measurements of action potential duration, beat period, spike amplitude, conduction velocity, and peak calcium-transient amplitude did not differ significantly between the two hiPSC-CM populations. Yet, our cardiac-origin iPSC-CM showed higher ADP and conduction velocity than previously reported iPSC-CM derived from non-cardiac tissues. Transcriptomic data comparing iPSC and iPSC-CMs showed similar gene expression profiles between AiPSC-CMs and ViPSC-CMs with significant differences when compared to iPSC-CM derived from other tissues. This analysis also pointed to several genes involved in electrophysiology processes to be responsible for the physiological differences observed between cardiac and non-cardiac-derived cardiomyocytes. ¬ Conclusions: AiPSC and ViPSC were differentiated into CMs with equal efficiency. Detected differences in electrophysiological properties, calcium handling activity, and transcription profiles between cardiac and non-cardiac derived cardiomyocytes demonstrated that 1) tissue of origin matters to generate a better-featured iPSC-CMs, 2) the sublocation within the cardiac tissue has marginal effects on the differentiation process.
2023-02-03 | GSE221268 | GEO
Project description:Evaluation of Manual and Automatic prepared HeliScopeCAGE cDNA
| PRJDB2160 | ENA
Project description:Engineered human cardiac tissues have been utilized for various biomedical applications, including drug testing, disease modeling, and regenerative medicine. However, the applications of cardiac tissues derived from human pluripotent stem cells are often limited due to their immaturity and lack of functionality. Therefore, in this study, we established a perfusable culture system based on in vivo-like heart microenvironments to improve human cardiac tissue fabrication. The integrated culture platform of a microfluidic chip and a three-dimensional heart extracellular matrix enhanced human cardiac tissue development and their structural and functional maturation. These tissues were comprised of cardiovascular lineage cells, including cardiomyocytes and cardiac fibroblasts derived from human induced pluripotent stem cells, as well as vascular endothelial cells. The resultant macroscale human cardiac tissues exhibited improved efficacy in drug testing (small molecules with various levels of arrhythmia risk), disease modeling (long QT syndrome and cardiac fibrosis), and regenerative therapy (myocardial infarction treatment). Therefore, our culture system can serve as a highly effective tissue-engineering platform to provide human cardiac tissues for versatile biomedical applications.
2024-02-03 | PXD042077 | Pride
Project description:Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have wide potential application in basic research, drug discovery, and regenerative medicine, but functional maturation remains challenging. Here, we present a simple method whereby maturation of hiPSC-CMs can be accelerated by simultaneous application of physiological Ca 2+ and frequency-ramped electrical pacing in culture. This combination produces realistic force-frequency relationship, physiological twitch kinetics, robust β-adrenergic response, improved Ca 2+ handling, and cardiac troponin I expression within 25 days. This study provides insights into the role of Ca 2+ in hiPSC-CM maturation and offers a scalable platform for translational and clinical research.
2022-04-28 | GSE201437 | GEO
Project description:Extracellular vesicles (EV) in body fluids are extensively studied as potential biomarkers for numerous diseases. Major impediments of EV-based biomarker discovery include the manual labor, and the specificity and reproducibility of EV sample preparation. To tackle this, we present an automated liquid handling workstation for the density-based separation of EV from human body fluids and compare its performance to manual handling by (in)experienced researchers. To validate automated density-based separation of EVs from human body fluids, including blood plasma and urine, we assess variation, yield and purity by mass spectrometry-based proteomics.
2023-05-10 | PXD032394 | Pride
Project description:Human cardiomyocytes can be generated from human embryonic stem cells (hESCs) in vitro by a variety of methods, including co-culture with visceral endoderm-like cell lines and growth factor directed differentiation as monolayers or three-dimensional embryonic bodies. To enable the identification, purification and characterisation of human embryonic stem cell derived cardiomyocytes (CMs) and cardiac progenitor cells (CPCs), we introduced sequences encoding GFP into the NKX2-5 locus by homologous recombination. We found that NKX2-5GFP hESCs facilitate quantification of cardiac differentiation, purification of hESC-derived committed cardiac progenitor cells and cardiomyocytes and the standardization of differentiation protocols.
2011-11-16 | E-MEXP-3371 | biostudies-arrayexpress