Journal ArticleDOI
Differentiation of cardiomyocytes and generation of human engineered heart tissue
K. Breckwoldt,David Letuffe-Brenière,Ingra Mannhardt,Thomas G. Schulze,Bärbel M. Ulmer,Tessa Werner,Anika Benzin,Birgit Klampe,Marina Reinsch,Sandra D. Laufer,Aya Shibamiya,Maksymilian Prondzynski,Giulia Mearini,Dennis Schade,Sigrid Fuchs,Christiane Neuber,Elisabeth Krämer,Umber Saleem,Mirja L Schulze,Marita L. Rodriguez,Thomas Eschenhagen,Arne Hansen +21 more
TLDR
This protocol describes the differentiation of cardiomyocytes from hiPSCs; standardized generation of defined embryoid bodies, growth factor and small-molecule-based cardiac differentiation; and standardizedgeneration of EHTs.Abstract:
Since the advent of the generation of human induced pluripotent stem cells (hiPSCs), numerous protocols have been developed to differentiate hiPSCs into cardiomyocytes and then subsequently assess their ability to recapitulate the properties of adult human cardiomyocytes. However, hiPSC-derived cardiomyocytes (hiPSC-CMs) are often assessed in single-cell assays. A shortcoming of these assays is the limited ability to characterize the physiological parameters of cardiomyocytes, such as contractile force, due to random orientations. This protocol describes the differentiation of cardiomyocytes from hiPSCs, which occurs within 14 d. After casting, cardiomyocytes undergo 3D assembly. This produces fibrin-based engineered heart tissues (EHTs)-in a strip format-that generate force under auxotonic stretch conditions. 10-15 d after casting, the EHTs can be used for contractility measurements. This protocol describes parallel expansion of hiPSCs; standardized generation of defined embryoid bodies, growth factor and small-molecule-based cardiac differentiation; and standardized generation of EHTs. To carry out the protocol, experience in advanced cell culture techniques is required.read more
Citations
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Journal ArticleDOI
Biomanufacturing of organ-specific tissues with high cellular density and embedded vascular channels
Mark A. Skylar-Scott,Mark A. Skylar-Scott,Sebastien G. M. Uzel,Sebastien G. M. Uzel,Lucy Nam,Lucy Nam,John H. Ahrens,John H. Ahrens,Ryan L. Truby,Ryan L. Truby,Sarita Damaraju,Sarita Damaraju,Jennifer A. Lewis,Jennifer A. Lewis +13 more
TL;DR: This work reports a biomanufacturing method for assembling hundreds of thousands of stem cell–derived organ building blocks into living matrices with high cellular density into which perfusable vascular channels are introduced via embedded three-dimensional bioprinting.
Patent
Differentiation of human embryonic stem cells
TL;DR: In this paper, a method for promoting the differentiation of pluripotent stem cells into insulin-producing cells was proposed, which is more than 80% of the cells within the population expressing characteristic markers of endodermal complete system.
Journal ArticleDOI
SARS-CoV-2 infects and induces cytotoxic effects in human cardiomyocytes.
Denisa Bojkova,Julian U. G. Wagner,Mariana Shumliakivska,Galip Servet Aslan,Umber Saleem,Arne Hansen,Guillermo Luxán,Stefan Günther,Minh Duc Pham,Jaya Krishnan,Patrick N. Harter,Utz H Ermel,Achilleas S. Frangakis,Hendrik Milting,Andreas M. Zeiher,Andreas M. Zeiher,Karin Klingel,Jindrich Cinatl,Andreas Dendorfer,Andreas Dendorfer,Thomas Eschenhagen,Carsten Tschöpe,Sandra Ciesek,Sandra Ciesek,Stefanie Dimmeler +24 more
TL;DR: It is demonstrated that SARS-CoV-2 infects cardiomyocytes in vitro in an ACE2- and cathepsin-dependent manner and is inhibited by the antiviral drug remdesivir, which warrants further monitoring of cardiotoxic effects in COVID-19 patients.
Journal ArticleDOI
Metabolic Maturation Media Improve Physiological Function of Human iPSC-Derived Cardiomyocytes
Dries A. M. Feyen,Dries A. M. Feyen,Wesley L. McKeithan,Wesley L. McKeithan,Wesley L. McKeithan,Arne A. N. Bruyneel,Sean Spiering,Sean Spiering,Larissa Hörmann,Bärbel M. Ulmer,Hui Zhang,Francesca Briganti,Michaela Schweizer,Bence Hegyi,Zhandi Liao,Risto Pekka Pölönen,Kenneth S. Ginsburg,Chi Keung Lam,Ricardo Serrano,Christine Wahlquist,Christine Wahlquist,Christine Wahlquist,Alexander Kreymerman,Michelle Vu,Prashila L Amatya,Charlotta Sophie Behrens,Sara Ranjbarvaziri,Renee G.C. Maas,Renee G.C. Maas,Matthew Greenhaw,Daniel Bernstein,Joseph C. Wu,Donald M. Bers,Thomas Eschenhagen,Christian M. Metallo,Mark Mercola,Mark Mercola,Mark Mercola +37 more
TL;DR: The maturation media designed to provide oxidative substrates adapted to the metabolic needs of human iPSC (hiPSC)-CMs should increase the fidelity of hiPSC-CMs as disease models.
Journal ArticleDOI
Stem Cell Therapy in Heart Diseases – Cell Types, Mechanisms and Improvement Strategies
TL;DR: A large number of clinical trials have shown stem cell therapy to be a promising therapeutic approach for the treatment of cardiovascular diseases, but results obtained from clinical studies are inconsistent and stem cell-based improvement of heart performance and cardiac remodeling was found to be quite limited.
References
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Journal ArticleDOI
Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors
Kazutoshi Takahashi,Koji Tanabe,Mari Ohnuki,Megumi Narita,Tomoko Ichisaka,Kiichiro Tomoda,Shinya Yamanaka +6 more
TL;DR: It is demonstrated that iPS cells can be generated from adult human fibroblasts with the same four factors: Oct3/4, Sox2, Klf4, and c-Myc.
Journal ArticleDOI
Embryonic Stem Cell Lines Derived from Human Blastocysts
James A. Thomson,Joseph Itskovitz-Eldor,Sander S. Shapiro,Michelle A. Waknitz,Swiergiel Jennifer J,Vivienne S. Marshall,Jeffrey M. Jones +6 more
TL;DR: Human blastocyst-derived, pluripotent cell lines are described that have normal karyotypes, express high levels of telomerase activity, and express cell surface markers that characterize primate embryonic stem cells but do not characterize other early lineages.
Journal ArticleDOI
Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors
Journal ArticleDOI
Cardiomyocytes derived from human embryonic stem cells in pro-survival factors enhance function of infarcted rat hearts
Michael A. Laflamme,Kent Chen,Anna V. Naumova,Veronica Muskheli,James A. Fugate,Sarah K. Dupras,Hans Reinecke,Chunhui Xu,Mohammad Hassanipour,Chris O'Sullivan,Lila R. Collins,Yinhong Chen,Elina Minami,Edward A. Gill,Shuichi Ueno,Chun Yuan,Joseph K. Gold,Charles E. Murry +17 more
TL;DR: This work generated highly purified human cardiomyocytes using a readily scalable system for directed differentiation that relies on activin A and BMP4, and identified a cocktail of pro-survival factors that limitsCardiomyocyte death after transplantation.
Journal ArticleDOI
Feeder-free growth of undifferentiated human embryonic stem cells.
Chunhui Xu,Margaret S. Inokuma,Jerrod Denham,Kathaleen Golds,Pratima Kundu,Joseph D. Gold,Melissa K. Carpenter +6 more
TL;DR: A successful feeder-free hES culture system in which undifferentiated cells can be maintained for at least 130 population doublings and are suitable for scaleup production is demonstrated.