scholarly journals Let-7 family of microRNA is required for maturation and adult-like metabolism in stem cell-derived cardiomyocytes

2015 ◽  
Vol 112 (21) ◽  
pp. E2785-E2794 ◽  
Author(s):  
Kavitha T. Kuppusamy ◽  
Daniel C. Jones ◽  
Henrik Sperber ◽  
Anup Madan ◽  
Karin A. Fischer ◽  
...  
Keyword(s):  
Stem Cell ◽  
Fatty Acid ◽  
Large Scale ◽  
Metabolic Flux ◽  
Embryonic Stem ◽  
Acid Oxidation ◽  
Loss Of Function ◽  
Metabolic Switch ◽  
Cardiac Maturation

In metazoans, transition from fetal to adult heart is accompanied by a switch in energy metabolism-glycolysis to fatty acid oxidation. The molecular factors regulating this metabolic switch remain largely unexplored. We first demonstrate that the molecular signatures in 1-year (y) matured human embryonic stem cell-derived cardiomyocytes (hESC-CMs) are similar to those seen in in vivo-derived mature cardiac tissues, thus making them an excellent model to study human cardiac maturation. We further show that let-7 is the most highly up-regulated microRNA (miRNA) family during in vitro human cardiac maturation. Gain- and loss-of-function analyses of let-7g in hESC-CMs demonstrate it is both required and sufficient for maturation, but not for early differentiation of CMs. Overexpression of let-7 family members in hESC-CMs enhances cell size, sarcomere length, force of contraction, and respiratory capacity. Interestingly, large-scale expression data, target analysis, and metabolic flux assays suggest this let-7–driven CM maturation could be a result of down-regulation of the phosphoinositide 3 kinase (PI3K)/AKT protein kinase/insulin pathway and an up-regulation of fatty acid metabolism. These results indicate let-7 is an important mediator in augmenting metabolic energetics in maturing CMs. Promoting maturation of hESC-CMs with let-7 overexpression will be highly significant for basic and applied research.

scholarly journals Lipid availability influences the metabolic maturation of human pluripotent stem cell-derived cardiomyocytes

2020 ◽  
Author(s):  
Hui Zhang ◽  
Mehmet G. Badur ◽  
Sean Spiering ◽  
Ajit Divakaruni ◽  
Noah E. Meurs ◽  
...  
Keyword(s):  
Stem Cell ◽  
Fatty Acid ◽  
Fatty Acid Oxidation ◽  
Pluripotent Stem Cell ◽  
Metabolic Flux ◽  
Acid Oxidation ◽  
Flux Analysis ◽  
Human Pluripotent Stem Cell

AbstractObjectivesPluripotent stem cell-derived cardiomyocytes are phenotypically immature, which limits their utility in downstream applications. Metabolism is dramatically reprogramed during cardiac maturation in vivo and presents a potential avenue to drive in vitro maturation. We aimed to identify and address metabolic bottlenecks in the generation of human pluripotent stem cell (hPSC)-derived cardiomyocytes.MethodshPSCs were differentiated into cardiomyocytes using an established, chemically-defined differentiation protocol. We applied 13C metabolic flux analysis (MFA) and targeted transcriptomics to characterize cardiomyocyte metabolism in during differentiation in the presence or absence of exogenous lipids.ResultshPSC-derived cardiomyocytes induced some cardiometabolic pathways (i.e. ketone body and branched-chain amino acid oxidation) but failed to effectively activate fatty acid oxidation. MFA studies indicated that lipid availability in cultures became limited during differentiation, suggesting potential issues with nutrient availability. Exogenous supplementation of lipids improved cardiomyocyte morphology, mitochondrial function, and promoted increased fatty acid oxidation in hPSC-derivatives.ConclusionhPSC-derived cardiomyocytes are dependent upon exogenous sources of lipids for metabolic maturation. Proper supplementation removes a potential roadblock in the generation of metabolically mature cardiomyocytes. These studies further highlight the importance of considering and exploiting metabolic phenotypes in the in vitro production and utilization of functional hPSC-derivatives.

The Ski oncoprotein regulates Hematopoietic Stem Cell Fitness and Functions as a Corepressor for Gfi1.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1403-1403
Author(s):  
Chinavenmeni S. Velu ◽  
Michael Berk ◽  
Haiming Xu ◽  
Tristan Bourdeau ◽  
Avedis Kazanjian ◽  
...  
Keyword(s):  
Stem Cell ◽  
Hematopoietic Stem Cell ◽  
Target Genes ◽  
Fetal Liver ◽  
Embryonic Stem ◽  
Loss Of Function ◽  
Hematopoietic Stem ◽  
Myeloid Progenitor ◽  
Multipotent Progenitors

Abstract Ski is a corepressor protein originally identified as a retrovirally transduced oncoprotein. Genetic deletion of Ski has revealed essential roles in multiple developmental processes. Suggestion that Ski may play a role in hematopoiesis first came from expression of v-Ski and c-Kit, which induced the continuous in vitro growth of primary avian multipotent progenitors. However, the hematopoietic phenotype of Ski−/− mice has not been described. Here, we show that Ski loss of function results in loss of hematopoietic stem cell (HSC) fitness and abnormal regulation of myeloid progenitor numbers. Fetal liver Ski−/− HSC engraft well in ablated recipients, but are not competitive in engraftment. Moreover, Ski null embryonic stem cells generate many tissues in chimeras, but infrequently participate in hematopoiesis. Thus, Ski null HSC are not competitive in both transplant and chimera settings, indicating a defect in stem cell fitness. Engrafted Ski−/− fetal liver cells generate fewer myeloid lineage cells than wild type littermates, and accumulate granulocytemonocyte progenitors. Growth factor independent -1 (Gfi1) is a transcriptional repressor that controls HSC maintenance and myeloid progenitor differentiation. Gfi1−/− and Ski−/− hematopoietic stem and myeloid progenitor phenotypes are strikingly similar. We find that Ski functions as a corepressor for Gfi1. Both endogenous and synthetic Gfi1 and Ski physically interact in vitro and upon Gfi1 target genes. Knockdown of Gfi1 or Ski results in derepression of these targets. Thus, our results provide a molecular link between the similar HSC and myeloid progenitor phenotypes engendered by Gfi1 or Ski deletion.

scholarly journals Potential Clinical Applications for Human Pluripotent Stem Cell-Derived Blood Components

2011 ◽  
Vol 2011 ◽  
pp. 1-11 ◽  
Author(s):  
Erin A. Kimbrel ◽  
Shi-Jiang Lu
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Large Scale ◽  
Embryonic Stem ◽  
Adult Stem Cell ◽  
Cell Types ◽  
The Body ◽  
Blood Components ◽  
Induced Pluripotent

The ability of human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) to divide indefinitely without losing pluripotency and to theoretically differentiate into any cell type in the body makes them highly attractive cell sources for large scale regenerative medicine purposes. The current use of adult stem cell-derived products in hematologic intervention sets an important precedent and provides a guide for developing hESC/iPSC based therapies for the blood system. In this review, we highlight biological functions of mature cells of the blood, clinical conditions requiring the transfusion or stimulation of these cells, and the potential for hESC/iPSC-derivatives to serve as functional replacements. Many researchers have already been able to differentiate hESCs and/or iPSCs into specific mature blood cell types. For example, hESC-derived red blood cells and platelets are functional in tasks such as oxygen delivery and blood clotting, respectively and may be able to serve as substitutes for their donor-derived counterparts in emergencies. hESC-derived dendritic cells are functional in antigen-presentation and may be used as off-the-shelf vaccine therapies to stimulate antigen-specific immune responses against cancer cells. However,in vitrodifferentiation systems used to generate these cells will need further optimization before hESC/iPSC-derived blood components can be used clinically.

scholarly journals Physiological and pharmacological stimulation for in vitro maturation of substrate metabolism in human induced pluripotent stem cell-derived cardiomyocytes

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Colleen A. Lopez ◽  
Heba Hussain A. A. Al-Siddiqi ◽  
Ujang Purnama ◽  
Sonia Iftekhar ◽  
Arne A. N. Bruyneel ◽  
...  
Keyword(s):  
Stem Cell ◽  
Fatty Acid ◽  
Fatty Acid Oxidation ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Acid Oxidation ◽  
Substrate Metabolism ◽  
Pharmacological Stimulation ◽  
Induced Pluripotent

AbstractHuman induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) enable human cardiac cells to be studied in vitro, although they use glucose as their primary metabolic substrate and do not recapitulate the properties of adult cardiomyocytes. Here, we have explored the interplay between maturation by stimulation of fatty acid oxidation and by culture in 3D. We have investigated substrate metabolism in hiPSC-CMs grown as a monolayer and in 3D, in porous collagen-derived scaffolds and in engineered heart tissue (EHT), by measuring rates of glycolysis and glucose and fatty acid oxidation (FAO), and changes in gene expression and mitochondrial oxygen consumption. FAO was stimulated by activation of peroxisome proliferator-activated receptor alpha (PPARα), using oleate and the agonist WY-14643, which induced an increase in FAO in monolayer hiPSC-CMs. hiPSC-CMs grown in 3D on collagen-derived scaffolds showed reduced glycolysis and increased FAO compared with monolayer cells. Activation of PPARα further increased FAO in cells on collagen/elastin scaffolds but not collagen or collagen/chondroitin-4-sulphate scaffolds. In EHT, FAO was significantly higher than in monolayer cells or those on static scaffolds and could be further increased by culture with oleate and WY-14643. In conclusion, a more mature metabolic phenotype can be induced by culture in 3D and FAO can be incremented by pharmacological stimulation.

scholarly journals PPARdelta signaling activation improves metabolic and contractile maturation of human pluripotent stem cell-derived cardiomyocytes.

2021 ◽  
Author(s):  
Nadeera M Wickramasinghe ◽  
David Sachs ◽  
Bhavana Shewale ◽  
David M Gonzalez ◽  
Priyanka Dhanan-Krishnan ◽  
...  
Keyword(s):  
Stem Cell ◽  
Heart Development ◽  
Regulatory Networks ◽  
Pluripotent Stem Cell ◽  
Disease Modeling ◽  
Acid Oxidation ◽  
Metabolic Switch ◽  
Ppar Signaling ◽  
Signaling Activation ◽  
Cardiac Maturation

Pluripotent stem cell-derived cardiomyocytes (PSC-CMs) provide an unprecedented opportunity to study human heart development and disease. A major caveat however is that they remain functionally and structurally immature in culture, limiting their potential for disease modeling and regenerative approaches. Here, we address the question of how different metabolic pathways can be modulated in order to induce efficient hPSC-CM maturation. We show that PPAR signaling acts in an isoform-specific manner to balance glycolysis and fatty acid oxidation (FAO). PPARD activation or inhibition results in efficient respective up- or down-regulation of the gene regulatory networks underlying FAO in hPSC-CMs. PPARD induction further increases mitochondrial and peroxisome content, enhances mitochondrial cristae formation and augments FAO flux. Lastly PPARD activation results in enhanced myofibril organization and improved contractility. Transient lactate exposure, commonly used in hPSC-CM purification protocols, induces an independent program of cardiac maturation, but when combined with PPARD activation equally results in a metabolic switch to FAO. In summary, we identify multiple axes of metabolic modifications of hPSC-CMs and a role for PPARD signaling in inducing the metabolic switch to FAO in hPSC-CMs. Our findings provide new and easily implemented opportunities to generate mature hPSC-CMs for disease modeling and regenerative therapy.

scholarly journals Challenges of stem cell application in research and clinical practice – an update

2021 ◽  
Vol 9 (4) ◽  
pp. 160-164
Author(s):  
Maurycy Jankowski ◽  
Marie Machatkova ◽  
Pavel Ventruba ◽  
Elena Kistanova ◽  
Alexander Makarevich ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Clinical Practice ◽  
Large Scale ◽  
Adult Stem Cells ◽  
Embryonic Stem ◽  
Stem Cell Differentiation ◽  
Human Organism ◽  
Adverse Side Effect

Abstract There are multiple possible applications of stem cells in medicine, from cell-based therapies for degenerative and dystrophic conditions, through novel approaches in cancer treatment, to in vitro organ printing. However, there are still several challenges that need to be overcame before stem cells therapies can be successfully introduced worldwide on a large scale. These include sourcing of stem cells, preventing their aberrant progression and ethical concerns regarding their use in animals and humans. Among the multiple stem cell types present in the human organism from the period of embryonic development to adulthood, this review focuses on the three types that gain the most attention in relation to modern research: embryonic stem cells, induced pluripotent stem cells and adult stem cells. There are a number of obstacles that need to be removed before these cells can be widely applied in clinical practice, including the choice of the perfect source of stem cells, full elucidation of the mechanisms of stem cell differentiation and plasticity, and minimization of adverse side effect potential. Nonetheless, the focus of the scientific community on the topic of stem cells remains unhindered, bringing hope that all of the possible concerns will be addressed in the future.

scholarly journals The TGFβ Family in Human Placental Development at the Fetal-Maternal Interface

Biomolecules ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 453
Author(s):  
Susana M. Chuva de Sousa Lopes ◽  
Marta S. Alexdottir ◽  
Gudrun Valdimarsdottir
Keyword(s):  
Stem Cell ◽  
Transforming Growth Factor Beta ◽  
Molecular Mechanisms ◽  
Transforming Growth Factor ◽  
Cell Model ◽  
Embryonic Stem ◽  
Model Systems ◽  
Special Focus ◽  
Placental Development

Emerging data suggest that a trophoblast stem cell (TSC) population exists in the early human placenta. However, in vitro stem cell culture models are still in development and it remains under debate how well they reflect primary trophoblast (TB) cells. The absence of robust protocols to generate TSCs from humans has resulted in limited knowledge of the molecular mechanisms that regulate human placental development and TB lineage specification when compared to other human embryonic stem cells (hESCs). As placentation in mouse and human differ considerably, it is only with the development of human-based disease models using TSCs that we will be able to understand the various diseases caused by abnormal placentation in humans, such as preeclampsia. In this review, we summarize the knowledge on normal human placental development, the placental disease preeclampsia, and current stem cell model systems used to mimic TB differentiation. A special focus is given to the transforming growth factor-beta (TGFβ) family as it has been shown that the TGFβ family has an important role in human placental development and disease.

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