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Nathan J Palpant, Lil Pabon, Clayton E Friedman, Meredith Roberts, Brandon Hadland, Rebecca J Zaunbrecher, Irwin Bernstein, Ying Zheng, Charles E Murry
Human pluripotent stem cells (hPSCs) provide a valuable model for the study of human development and a means to generate a scalable source of cells for therapeutic applications. This protocol specifies cell fate efficiently into cardiac and endothelial lineages from hPSCs. The protocol takes 2 weeks to complete and requires experience in hPSC culture and differentiation techniques. Building on lessons taken from early development, this monolayer-directed differentiation protocol uses different concentrations of activin A and bone morphogenetic protein 4 (BMP4) to polarize cells into mesodermal subtypes that reflect mid-primitive-streak cardiogenic mesoderm and posterior-primitive-streak hemogenic mesoderm...
January 2017: Nature Protocols
Jia-Ling Ruan, Nathaniel L Tulloch, Maria V Razumova, Mark Saiget, Veronica Muskheli, Lil Pabon, Hans Reinecke, Michael Regnier, Charles E Murry
BACKGROUND: Tissue engineering enables the generation of functional human cardiac tissue with cells derived in vitro in combination with biocompatible materials. Human-induced pluripotent stem cell-derived cardiomyocytes provide a cell source for cardiac tissue engineering; however, their immaturity limits their potential applications. Here we sought to study the effect of mechanical conditioning and electric pacing on the maturation of human-induced pluripotent stem cell-derived cardiac tissues...
November 15, 2016: Circulation
Wan Qin, Meredith A Roberts, Xiaoli Qi, Charles E Murry, Ying Zheng, Ruikang K Wang
In this study, we propose a novel implementation of optical coherence tomography-based angiography combined with ex vivo perfusion of fixed hearts to visualize coronary microvascular structure and function. The extracorporeal perfusion of Intralipid solution allows depth-resolved angiographic imaging, control of perfusion pressure, and high-resolution optical microangiography. The imaging technique offers new opportunities for microcirculation research in the heart, which has been challenging due to motion artifacts and the lack of independent control of pressure and flow...
November 7, 2016: Physics in Medicine and Biology
Kevin S Bielawski, Andrea Leonard, Shiv Bhandari, Chuck E Murry, Nathan J Sniadecki
Engineered heart tissues made from human pluripotent stem cell-derived cardiomyocytes have been used for modeling cardiac pathologies, screening new therapeutics, and providing replacement cardiac tissue. Current methods measure the functional performance of engineered heart tissue by their twitch force and beating frequency, typically obtained by optical measurements. In this article, we describe a novel method for assessing twitch force and beating frequency of engineered heart tissue using magnetic field sensing, which enables multiple tissues to be measured simultaneously...
September 28, 2016: Tissue Engineering. Part C, Methods
Lindsey A Muir, Charles E Murry, Jeffrey S Chamberlain
In Duchenne muscular dystrophy (DMD) and other muscle wasting disorders, cell therapies are a promising route for promoting muscle regeneration by supplying a functional copy of the missing dystrophin gene and contributing new muscle fibers. The clinical application of cell-based therapies is resource intensive, and it will therefore be necessary to address key limitations that reduce cell engraftment into muscle tissue. A pressing issue is poor donor cell survival following transplantation, which in preclinical studies limits the ability to effectively test the impact of cell-based therapy on whole muscle function...
September 7, 2016: Stem Cells and Development
Brenda M Ogle, Nenad Bursac, Ibrahim Domian, Ngan F Huang, Philippe Menasché, Charles E Murry, Beth Pruitt, Milica Radisic, Joseph C Wu, Sean M Wu, Jianyi Zhang, Wolfram-Hubertus Zimmermann, Gordana Vunjak-Novakovic
The promise of cardiac tissue engineering is in the ability to recapitulate in vitro the functional aspects of a healthy heart and disease pathology as well as to design replacement muscle for clinical therapy. Parts of this promise have been realized; others have not. In a meeting of scientists in this field, five central challenges or "big questions" were articulated that, if addressed, could substantially advance the current state of the art in modeling heart disease and realizing heart repair.
June 8, 2016: Science Translational Medicine
Jonathan Kimmelman, Insoo Hyun, Nissim Benvenisty, Timothy Caulfield, Helen E Heslop, Charles E Murry, Douglas Sipp, Lorenz Studer, Jeremy Sugarman, George Q Daley
No abstract text is available yet for this article.
May 12, 2016: Nature
George Q Daley, Insoo Hyun, Jane F Apperley, Roger A Barker, Nissim Benvenisty, Annelien L Bredenoord, Christopher K Breuer, Timothy Caulfield, Marcelle I Cedars, Joyce Frey-Vasconcells, Helen E Heslop, Ying Jin, Richard T Lee, Christopher McCabe, Megan Munsie, Charles E Murry, Steven Piantadosi, Mahendra Rao, Heather M Rooke, Douglas Sipp, Lorenz Studer, Jeremy Sugarman, Masayo Takahashi, Mark Zimmerman, Jonathan Kimmelman
The International Society for Stem Cell Research (ISSCR) presents its 2016 Guidelines for Stem Cell Research and Clinical Translation (ISSCR, 2016). The 2016 guidelines reflect the revision and extension of two past sets of guidelines (ISSCR, 2006; ISSCR, 2008) to address new and emerging areas of stem cell discovery and application and evolving ethical, social, and policy challenges. These guidelines provide an integrated set of principles and best practices to drive progress in basic, translational, and clinical research...
June 14, 2016: Stem Cell Reports
Jonathan Kimmelman, Helen E Heslop, Jeremy Sugarman, Lorenz Studer, Nissim Benvenisty, Timothy Caulfield, Insoo Hyun, Charles E Murry, Douglas Sipp, George Q Daley
No abstract text is available yet for this article.
May 14, 2016: Lancet
Timothy Caulfield, Douglas Sipp, Charles E Murry, George Q Daley, Jonathan Kimmelman
No abstract text is available yet for this article.
May 13, 2016: Science
Josè Manuel Pioner, Alice W Racca, Jordan M Klaiman, Kai-Chun Yang, Xuan Guan, Lil Pabon, Veronica Muskheli, Rebecca Zaunbrecher, Jesse Macadangdang, Mark Y Jeong, David L Mack, Martin K Childers, Deok-Ho Kim, Chiara Tesi, Corrado Poggesi, Charles E Murry, Michael Regnier
Tension production and contractile properties are poorly characterized aspects of excitation-contraction coupling of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Previous approaches have been limited due to the small size and structural immaturity of early-stage hiPSC-CMs. We developed a substrate nanopatterning approach to produce hiPSC-CMs in culture with adult-like dimensions, T-tubule-like structures, and aligned myofibrils. We then isolated myofibrils from hiPSC-CMs and measured the tension and kinetics of activation and relaxation using a custom-built apparatus with fast solution switching...
June 14, 2016: Stem Cell Reports
Astrid Breitbart, Charles E Murry
There is still no curative treatment for Duchenne muscular dystrophy (DMD). In this issue of Cell Stem Cell, Young et al. (2016) demonstrate a genome editing approach applicable to 60% of DMD patients with CRISPR/Cas9 using one pair of guide RNAs.
April 7, 2016: Cell Stem Cell
Meredith A Roberts, Dominic Tran, Kareen L K Coulombe, Maria Razumova, Michael Regnier, Charles E Murry, Ying Zheng
Cardiac tissue engineering is a strategy to replace damaged contractile tissue and model cardiac diseases to discover therapies. Current cardiac and vascular engineering approaches independently create aligned contractile tissue or perfusable vasculature, but a combined vascularized cardiac tissue remains to be achieved. Here, we sought to incorporate a patterned microvasculature into engineered heart tissue, which balances the competing demands from cardiomyocytes to contract the matrix versus the vascular lumens that need structural support...
April 2016: Tissue Engineering. Part A
Daniel Carson, Marketa Hnilova, Xiulan Yang, Cameron L Nemeth, Jonathan H Tsui, Alec S T Smith, Alex Jiao, Michael Regnier, Charles E Murry, Candan Tamerler, Deok-Ho Kim
Understanding the phenotypic development of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) is a prerequisite to advancing regenerative cardiac therapy, disease modeling, and drug screening applications. Lack of consistent hiPSC-CM in vitro data can be largely attributed to the inability of conventional culture methods to mimic the structural, biochemical, and mechanical aspects of the myocardial niche accurately. Here, we present a nanogrid culture array comprised of nanogrooved topographies, with groove widths ranging from 350 to 2000 nm, to study the effect of different nanoscale structures on the structural development of hiPSC-CMs in vitro...
August 31, 2016: ACS Applied Materials & Interfaces
Paul A Bunn, John D Minna, Alexander Augustyn, Adi F Gazdar, Youcef Ouadah, Mark A Krasnow, Anton Berns, Elisabeth Brambilla, Natasha Rekhtman, Pierre P Massion, Matthew Niederst, Martin Peifer, Jun Yokota, Ramaswamy Govindan, John T Poirier, Lauren A Byers, Murry W Wynes, David G McFadden, David MacPherson, Christine L Hann, Anna F Farago, Caroline Dive, Beverly A Teicher, Craig D Peacock, Jane E Johnson, Melanie H Cobb, Hans-Guido Wendel, David Spigel, Julien Sage, Ping Yang, M Catherine Pietanza, Lee M Krug, John Heymach, Peter Ujhazy, Caicun Zhou, Koichi Goto, Afshin Dowlati, Camilla Laulund Christensen, Keunchil Park, Lawrence H Einhorn, Martin J Edelman, Giuseppe Giaccone, David E Gerber, Ravi Salgia, Taofeek Owonikoko, Shakun Malik, Niki Karachaliou, David R Gandara, Ben J Slotman, Fiona Blackhall, Glenwood Goss, Roman Thomas, Charles M Rudin, Fred R Hirsch
No abstract text is available yet for this article.
April 2016: Journal of Thoracic Oncology
Sarah Fernandes, James J H Chong, Sharon L Paige, Mineo Iwata, Beverly Torok-Storb, Gordon Keller, Hans Reinecke, Charles E Murry
Cardiomyocytes derived from human embryonic stem cells (hESC-CMs) can improve the contractility of injured hearts.We hypothesized that mesodermal cardiovascular progenitors (hESC-CVPs), capable of generating vascular cells in addition to cardiomyocytes, would provide superior repair by contributing to multiple components of myocardium. We performed a head-to-head comparison of hESC-CMs and hESC-CVPs and compared these with the most commonly used clinical cell type, human bone marrow mononuclear cells (hBMMNCs)...
November 10, 2015: Stem Cell Reports
Stephen C Kolwicz, Guy L Odom, Sarah G Nowakowski, Farid Moussavi-Harami, Xiaolan Chen, Hans Reinecke, Stephen D Hauschka, Charles E Murry, Gregory G Mahairas, Michael Regnier
Impaired systolic function, resulting from acute injury or congenital defects, leads to cardiac complications and heart failure. Current therapies slow disease progression but do not rescue cardiac function. We previously reported that elevating the cellular 2 deoxy-ATP (dATP) pool in transgenic mice via increased expression of ribonucleotide reductase (RNR), the enzyme that catalyzes deoxy-nucleotide production, increases myosin-actin interaction and enhances cardiac muscle contractility. For the current studies, we initially injected wild-type mice retro-orbitally with a mixture of adeno-associated virus serotype-6 (rAAV6) containing a miniaturized cardiac-specific regulatory cassette (cTnT(455)) composed of enhancer and promotor portions of the human cardiac troponin T gene (TNNT2) ligated to rat cDNAs encoding either the Rrm1 or Rrm2 subunit...
February 2016: Molecular Therapy: the Journal of the American Society of Gene Therapy
Desy S Lee, Jyh-Hong Chen, David J Lundy, Chung-Hung Liu, Shiaw-Min Hwang, Lil Pabon, Ru-Chi Shieh, Chien-Chang Chen, Sheng-Nan Wu, Yu-Ting Yan, Sho-Tone Lee, Po-Min Chiang, Shu Chien, Charles E Murry, Patrick C H Hsieh
Pluripotent-cell-derived cardiomyocytes have great potential for use in research and medicine, but limitations in their maturity currently constrain their usefulness. Here, we report a method for improving features of maturation in murine and human embryonic-stem-cell-derived cardiomyocytes (m/hESC-CMs). We found that coculturing m/hESC-CMs with endothelial cells improves their maturity and upregulates several microRNAs. Delivering four of these microRNAs, miR-125b-5p, miR-199a-5p, miR-221, and miR-222 (miR-combo), to m/hESC-CMs resulted in improved sarcomere alignment and calcium handling, a more negative resting membrane potential, and increased expression of cardiomyocyte maturation markers...
September 29, 2015: Cell Reports
Kaytlyn A Gerbin, Xiulan Yang, Charles E Murry, Kareen L K Coulombe
Cardiac tissue engineering is a promising approach to provide large-scale tissues for transplantation to regenerate the heart after ischemic injury, however, integration with the host myocardium will be required to achieve electromechanical benefits. To test the ability of engineered heart tissues to electrically integrate with the host, 10 million human embryonic stem cell (hESC)-derived cardiomyocytes were used to form either scaffold-free tissue patches implanted on the epicardium or micro-tissue particles (~1000 cells/particle) delivered by intramyocardial injection into the left ventricular wall of the ischemia/reperfusion injured athymic rat heart...
2015: PloS One
Nathan J Palpant, Lil Pabon, Meredith Roberts, Brandon Hadland, Daniel Jones, Christina Jones, Randall T Moon, Walter L Ruzzo, Irwin Bernstein, Ying Zheng, Charles E Murry
During vertebrate development, mesodermal fate choices are regulated by interactions between morphogens such as activin/nodal, BMPs and Wnt/β-catenin that define anterior-posterior patterning and specify downstream derivatives including cardiomyocyte, endothelial and hematopoietic cells. We used human embryonic stem cells to explore how these pathways control mesodermal fate choices in vitro. Varying doses of activin A and BMP4 to mimic cytokine gradient polarization in the anterior-posterior axis of the embryo led to differential activity of Wnt/β-catenin signaling and specified distinct anterior-like (high activin/low BMP) and posterior-like (low activin/high BMP) mesodermal populations...
September 15, 2015: Development
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