RNA sequencing of hPSC-derived cardiac progenitors and endocardium
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ABSTRACT:
PROVIDER: PRJNA427332 | ENA |
REPOSITORIES: ENA
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Project description:We utilized a dual reporting hPSC line that identified cells expressing NKX2.5 and endothelal cells to characterize discrete milestones during cardiac and vascular differentiaiton. Comparing populations that express either or both reporters to human umbilical vein endothelial cells, we document a unique molecular phenotype in hPSC-derived endocardium that points toward an important role for Wnt signaling during vascular specification of cardiac progenitors.
2017-12-27 | GSE108465 | GEO
Project description:Neurons derived from human pluripotent stem cells (hPSCs) are a remarkable tool for modeling human neural development and diseases. However, it remains largely unknown whether the hPSC-derived neurons can be functionally coupled with their target tissues in vitro, which is essential for understanding inter-cellular physiology and further translational studies. Here, we demonstrate that hPSC-derived sympathetic neurons can be obtained from hPSCs and that the resulting neurons form physical and functional connections with cardiac muscle cells. By use of multiple hPSC reporter lines, we recapitulated human autonomic neuron development in vitro, and successfully isolated PHOX2B::eGFP+ neurons exhibiting sympathetic marker expression, electrophysiological properties, and norepinephrine secretion. With pharmacological and optogenetic manipulations, the PHOX2B::eGFP+ neurons controlled the beating rates of cardiomyocytes, and their physical interaction led to neuronal maturation. Our study lays a foundation for the specification of human sympathetic neurons and for the hPSC-based neuronal control of end organs in a dish. Using the four genetic reporter systems (OCT4::eGFP, SOX10::eGFP, ASCL1::eGFP, and PHOX2B::eGFP reporter hESC lines), we were able to purify discrete cell populations at four differentiation stages, recapitulating the sympathoadrenal differentiation process in vitro with purified and defined populations in four specific differentiation stages. We performed transcriptome analysis of OCT4::eGFP+ cells (3 biological replicates, representing undifferentiated hESCs), SOX10::eGFP+ cells (3 biological replicates, multi-potent neural crest), ASCL1::eGFP+ cells (3 biological replicates, putative sympathoadrenal progenitors), and PHOX2B::eGFP+ cells (2 biological replicates, putative sympathetic neuronal precursors).
2016-07-01 | E-GEOD-80689 | biostudies-arrayexpress
Project description:Directed differentiation of human induced pluripotent stem cells creates billions of patient-specific cells. However, the differentiated derivatives of hiPS cells are immature relative to adult counterparts. Proliferative progenitor cells in these protocols are also therapeutically promising, but progenitors with enhanced potential for maturation have not been discovered. We hypothesized that brief epigenetic and innate immune modulation with polyinosinc cytidilic acid (pIC) could create cardiac progenitors with enhanced later maturation. Progenitor transcriptomic analysis revealed increases in cardiac crescent-like proliferative notch signaling by Jagged1, and activated progenitors spontaneously gave rise to cardiomyocytes with enhanced maturity and conductive microtissue with arrhythmogenic resistance. Furthermore, activated cardiac progenitors improved survival in animals after myocardial infarction compared to untreated progenitors. Our data suggest that future organization and maturation of cardiomyocytes are impacted by earlier pre-cardiomyocyte developmental signaling, and that progenitors modifying these pathways have translational promise. The forward epigenetic and immune modulation approach here utilized may be applicable to other cell lineages as well.
2018-08-01 | GSE98941 | GEO
Project description:Cardiac fibroblasts (CFs) play critical roles in heart development, homeostasis, and disease. The limited availability of human CFs from native heart impedes investigations of CF biology and their role in disease. Human pluripotent stem cells (hPSCs) provide a highly renewable and genetically defined cell source, but efficient methods to generate CFs from hPSCs have not been described. Here, we show differentiation of hPSCs using sequential modulation of Wnt and FGF signaling to generate second heart field progenitors that efficiently give rise to hPSC-CFs. The hPSC-CFs resemble native heart CFs in cell morphology, proliferation, gene expression, fibroblast marker expression, production of extracellular matrix and myofibroblast transformation induced by TGFβ1 and angiotensin II. Furthermore, hPSC-CFs exhibit a more embryonic phenotype when compared to fetal and adult primary human CFs. Co-culture of hPSC-CFs with hPSC-derived cardiomyocytes distinctly alters the electrophysiological properties (EP) of the cardiomyocytes compared to co-culture with dermal fibroblasts (DFs). The hPSC-CFs provide a powerful cell source for research, drug discovery, precision medicine, and therapeutic applications in cardiac regeneration.
2019-02-08 | GSE126260 | GEO
Project description:In the heart, cardiomyocytes (CMs) are coupled to capillary endothelial cells (EC), mural cells (e.g. pericytes) and fibroblasts (Fb) promoting structural and electrophysiological tissue maturation as well as vascular network formation. Here, an in vitro model is shown for the investigation of the role of ECs, cardiac pericyte-like cells (PC) and different Fb sources in hPSC-derived bioartificial cardiac tissue (BCT) formation and function. The hPSC-based CMs, ECs, and PCs were differentiated, purified, and characterized for cell-type specific marker expression and function. Differentiated hPSC-PCs were used with hPSC-ECs to generate BCTs and to address their effect on tissue morphology and electromechanical parameters compared to control tissues containing primary dermal or cardiac Fbs.
2020-11-30 | GSE146150 | GEO
Project description:Neurons derived from human pluripotent stem cells (hPSCs) are a remarkable tool for modeling human neural development and diseases. However, it remains largely unknown whether the hPSC-derived neurons can be functionally coupled with their target tissues in vitro, which is essential for understanding inter-cellular physiology and further translational studies. Here, we demonstrate that hPSC-derived sympathetic neurons can be obtained from hPSCs and that the resulting neurons form physical and functional connections with cardiac muscle cells. By use of multiple hPSC reporter lines, we recapitulated human autonomic neuron development in vitro, and successfully isolated PHOX2B::eGFP+ neurons exhibiting sympathetic marker expression, electrophysiological properties, and norepinephrine secretion. With pharmacological and optogenetic manipulations, the PHOX2B::eGFP+ neurons controlled the beating rates of cardiomyocytes, and their physical interaction led to neuronal maturation. Our study lays a foundation for the specification of human sympathetic neurons and for the hPSC-based neuronal control of end organs in a dish.
2016-07-01 | GSE80689 | GEO
Project description:Despite the progress in safety and efficacy of cell therapy with pluripotent stem cells (PSCs), the presence of residual undifferentiated stem cells or proliferating neural progenitor cells (NPCs) with rostral identity has remained a major challenge. Here we reported the generation of an LMX1A knock-in GFP reporter human embryonic stem cell (hESC) line that marks the early dopaminergic progenitors during neural differentiation. Purified GFP positive cells in vitro exhibited expression of mRNA and proteins that characterized and matched the midbrain dopaminergic identity. Further proteomic analysis of enriched LMX1A+ cells identified several membrane associated proteins including CNTN2, enabling prospective isolation of LMX1A+ progenitor cells. Transplantation of hPSC-derived purified CNTN2+ progenitors enhanced dopamine release from transplanted cells in the host brain and alleviated Parkinson’s disease symptoms in animal models. Our study establishes an efficient approach for purification of large numbers of hPSC-derived dopaminergic progenitors for therapeutic applications.
2018-06-06 | PXD007837 | Pride
Project description:Mamamlian cardiogenesis occurs through the development of discreate populations of first and second heart field progenitors. We have used a dual transgenic color reproter system to isolate purified populations of these progenitors. We used microarrays to detail the global programme of gene expression underlying cardiac development in mouse; All four populations of cells are derived from embryonic stem cells differentiating in vitro (day 6 of in vitro differentaition). The stem cell line has two transgenic reporters as follows:; 1. The second heart field (SHF) specific reporter of the Mef2C gene (E. Dodou, S. M. Xu, B. L. Black, Mech Dev 120, 1021 (Sep, 2003)) driving the expression of dsRed; 2. The cardiac specific enhancer ( C. L. Lien et al., Development 126, 75 (Jan, 1999)) driving the expression of eGFP. Thus, the red cells are SHF specific, the green cells are cardiac specific, and the red/green are SHF and cardiac specific. These cells are compared to the double negative cells which serve as a control. Experiment Overall Design: Embryonic stem cell derived progenitors were isolated into four distinct populations by FACS purifying these progenitors based on a two color reporter system. Four populations were then compared to each other by transcriptional profiling.
2009-09-12 | E-GEOD-17513 | biostudies-arrayexpress
Project description:Pathogenic mutations in alpha kinase 3 (ALPK3) cause cardiomyopathy and a range of musculoskeletal defects. How ALPK3 mutations result in disease remains unclear and little is known about this atypical kinase. Using a suite of engineered human pluripotent stem cells (hPSCs) we show that ALPK3 localizes to the sarcomere, specifically at the M-Band. Both sarcomeric organization and calcium kinetics were disrupted in ALPK3 deficient hPSC derived cardiomyocytes. Further, cardiac organoids derived from ALPK3 knockout hPSCs displayed reduced force generation. Phosphoproteomic profiling of wildtype and ALPK3 null hPSC derived cardiomyocytes revealed ALPK3-dependant phospho-peptides were enriched for proteins involved in sarcomere function and protein quality control. We demonstrate that ALPK3 binds to the selective autophagy receptor SQSTM1 (Sequestome 1) and is required for the sarcomeric localization of SQSTM1. We propose that ALPK3 is a myogenic kinase with an integral role in the intracellular signaling networks underlying sarcomere maintenance required for continued cardiac contractility.
2023-01-19 | PXD035535 | Pride
Project description:Background and Objective: Currently, the cells for transplantation were derived from either autologous or allogeneic tissue. The former has a drawback that the quality of donor cells could depend on the patient’s condition, and the quantity could also be limited. To solve these problems, we investigated the potential of allogeneic cardiac mesenchymal progenitors (CMPs) derived from postmortem heart, which might be an immunological privileged like bone marrow-derived mesenchymal progenitors. Materials and Methods: We examined whether viable CMPs could be isolated from murine postmortem cardiac tissue that was harvested 24 hours postmortem. After two to three weeks propagation with high dose of basic fibroblast growth factor, we performed the cellular characteristics analyses, which included proliferation and differentiation property flow cytometric analyses, and microarray analyses. Results: Postmortem CMPs had longer lag phase after seeding than CMPs from living tissues, but they demonstrated the similar characteristics in all above examinations. In addition, global gene expression analysis by microarray indicated the similar characteristics between the cell derived from postmortem and living tissue. Conclusion: These results indicate allogeneic postmortem CMPs could have promising potential for cell transplantation as clinical applications, because of circumventing the issue of brain death. The samples were collected fom living or postmortem cardiac tissue (24 hr at 4C). We generated cardiac mesenchymal progenitors (CMPs) from these cardiac tissue, and compared global gene expression by AgilentMouse GE 8x60k Microarray. Adult, or fetal mouse heart RNAs were used as positive control. Adult mouse total heart RNAs were purchased from Clontech. Fetal mouse heart was extracted from fetus which is embryonic day 16.5 C57BL/6 strain. Tg means Transgenic mouse (C57BL/6-Tg(Myh6-EGFP)MG2).
2013-08-28 | E-GEOD-50221 | biostudies-arrayexpress