Name: suppression of pro-fibrotic signaling potentiates factor-mediated reprogramming of mouse embryonic fibroblasts into induced cardiomyocytes
Uploaded: 2018-06-03T14:00:00
Description: Watch this Scientific Journal Video about Suppression of Pro-fibrotic Signaling Potentiates Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts into Induced Cardiomyocytes at JoVE.com

Denna forskning stöddes av medel från Boettcher stiftelsens Webb-Waring biomedicinska forskningsprogram, amerikansk hjärtat Association vetenskapsman utveckling Grant (13SDG17400031), University of Colorado Institutionen för medicin utestående tidig karriär Scholar Program, University of Colorado avdelningen för kardiologi Barlow Nyle kapitalförsäkring och NIH R01HL133230 (till KS). A.S.R stöddes av NIH/NCATS Colorado CTSA Grant nummer TL1TR001081 och en pre doktorand stipendium från University of Colorado konsortiet för fibros forskning & översättning (CFReT). Denna forskning stöddes också av Cancer Center stöd beviljande (P30CA046934), den kärnor forskningsbidrag för hud sjukdomar (P30AR057212) och Flow flödescytometri kärnan vid University of Colorado Anschutz Medical Campus.

Alla experiment som kräver djur godkändes av institutionella djur vård och användning kommittén vid UC Denver Anschutz Medical Campus.
1. isolering av MEFs
<ol><li>Köpa C57BL/6 dräktiga möss på E13. Fartyg över natten.</li>
	<li>Euthanize mor enligt godkända IACUC protokoll (ex: ~1.3 L/min CO2 tills djuret visas döda följt av cervikal dislokation)</li>
	<li>Spray mor med 70% etanol och öppna bukhålan. Ta bort livmoder horn som innehåller embryon och placera i en 10...

<ol>
	<li>Benjamin, E. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heart+Disease+and+Stroke+Statistics-2017+Update:+A+Report+From+the+American+Heart+Association.">Heart Disease and Stroke Statistics-2017 Update: A Report From the American Heart Association.</a> Circulation. 136 (20), (2017).</li><li>Laflamme, M. A., Murry, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regenerating+the+heart.">Regenerating the heart.</a> Nat. Biotechnol. 23 (7), 845-856 (2005).</li><li>Mercola, M., Ruiz-Lozano, P., Schneider, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+muscle+regeneration:+lessons+from+development.">Cardiac muscle regeneration: lessons from development.</a> Genes &amp; Development. 25 (4), 299-309 (2011).</li><li>Bergmann, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+for+cardiomyocyte+renewal+in+humans.">Evidence for cardiomyocyte renewal in humans.</a> Science. 324 (5923), 98-102 (2009).</li><li>Senyo, S. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mammalian+heart+renewal+by+pre-existing+cardiomyocytes.">Mammalian heart renewal by pre-existing cardiomyocytes.</a> Nature. 493 (7432), 433-436 (2013).</li><li>Nabel, E. G., Braunwald, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+tale+of+coronary+artery+disease+and+myocardial+infarction.">A tale of coronary artery disease and myocardial infarction.</a> N Engl J Med. 366 (1), 54-63 (2012).</li><li>Packer, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+carvedilol+on+the+morbidity+of+patients+with+severe+chronic+heart+failure:+results+of+the+carvedilol+prospective+randomized+cumulative+survival+(COPERNICUS)+study.">Effect of carvedilol on the morbidity of patients with severe chronic heart failure: results of the carvedilol prospective randomized cumulative survival (COPERNICUS) study.</a> Circulation. 106 (17), 2194-2199 (2002).</li><li>The CAPRICORN Investigators. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+carvedilol+on+outcome+after+myocardial+infarction+in+patients+with+left-ventricular+dysfunction:+the+CAPRICORN+randomized+trial.">Effect of carvedilol on outcome after myocardial infarction in patients with left-ventricular dysfunction: the CAPRICORN randomized trial.</a> The Lancet. 357 (9266), 1385-1390 (2001).</li><li>Marangou, J., Paul, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+attitudes+on+cardiac+devices+in+heart+failure:+a+review.">Current attitudes on cardiac devices in heart failure: a review.</a> Clin Ther. 37 (10), 2206-2214 (2015).</li><li>Xin, M., Olson, E. N., Bassel-Duby, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mending+broken+hearts:+cardiac+development+as+a+basis+for+adult+heart+regeneration+and+repair.">Mending broken hearts: cardiac development as a basis for adult heart regeneration and repair.</a> Nat Rev Mol Cell Bio. 14 (8), 529-541 (2013).</li><li>Wollert, K. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracoronary+autologous+bone-marrow+cell+transfer+after+myocardial+infarction:+the+BOOST+randomized+controlled+clinical+trial.">Intracoronary autologous bone-marrow cell transfer after myocardial infarction: the BOOST randomized controlled clinical trial.</a> Lancet. 364 (9429), 141-148 (2004).</li><li>Schachinger, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracoronary+bone+marrow-derived+progenitor+cells+in+acute+myocardial+infarction.">Intracoronary bone marrow-derived progenitor cells in acute myocardial infarction.</a> N. Engl. J. Med. 355 (12), 1210-1221 (2006).</li><li>Huikuri, H. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+intracoronary+injection+of+mononuclear+bone+marrow+cells+on+left+ventricular+function,+arrhythmia+risk+profile,+and+restenosis+after+thrombolytic+therapy+of+acute+myocardial+infarction.">Effects of intracoronary injection of mononuclear bone marrow cells on left ventricular function, arrhythmia risk profile, and restenosis after thrombolytic therapy of acute myocardial infarction.</a> Eur. Heart J. 29 (22), 2723-2732 (2008).</li><li>Janssens, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autologous+bone+marrow-derived+stem-cell+transfer+in+patients+with+ST-segment+elevation+myocardial+infarction:+double-blind,+randomised+controlled+trial.">Autologous bone marrow-derived stem-cell transfer in patients with ST-segment elevation myocardial infarction: double-blind, randomised controlled trial.</a> Lancet. 367 (9505), 113-121 (2006).</li><li>Lunde, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracoronary+injection+of+mononuclear+bone+marrow+cells+in+acute+myocardial+infarction.">Intracoronary injection of mononuclear bone marrow cells in acute myocardial infarction.</a> N. Engl. J. Med. 355 (12), 1199-1209 (2006).</li><li>Hirsch, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracoronary+infusion+of+autologous+mononuclear+bone+marrow+cells+or+peripheral+mononuclear+blood+cells+after+primary+percutaneous+coronary+intervention:+rationale+and+design+of+the+HEBE+trial+-+a+prospective,+multicenter,+randomized+trial.">Intracoronary infusion of autologous mononuclear bone marrow cells or peripheral mononuclear blood cells after primary percutaneous coronary intervention: rationale and design of the HEBE trial - a prospective, multicenter, randomized trial.</a> Am. Heart J. 152 (3), 434-441 (2006).</li><li>Hirsch, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracoronary+infusion+of+mononuclear+cells+from+bone+marrow+or+peripheral+blood+compared+with+standard+therapy+in+patients+after+acute+myocardial+infarction+treated+by+primary+percutaneous+coronary+intervention:+results+of+the+randomized+controlled+HEBE+trial.">Intracoronary infusion of mononuclear cells from bone marrow or peripheral blood compared with standard therapy in patients after acute myocardial infarction treated by primary percutaneous coronary intervention: results of the randomized controlled HEBE trial.</a> Eur. Heart J. 32 (14), 1736-1747 (2011).</li><li>Karantalis, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autologous+mesenchymal+stem+cells+produce+concordant+improvements+in+regional+function,+tissue+perfusion,+and+fibrotic+burden+when+administered+to+patients+undergoing+coronary+artery+bypass+grafting:+The+Prospective+Randomized+Study+of+Mesenchymal+Stem+Cell+Therapy+in+Patients+Undergoing+Cardiac+Surgery+(PROMETHEUS)+trial.">Autologous mesenchymal stem cells produce concordant improvements in regional function, tissue perfusion, and fibrotic burden when administered to patients undergoing coronary artery bypass grafting: The Prospective Randomized Study of Mesenchymal Stem Cell Therapy in Patients Undergoing Cardiac Surgery (PROMETHEUS) trial.</a> Circ Res. 114 (8), 1302-1310 (2014).</li><li>Takahashi, K., Yamanaka, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+pluripotent+stem+cells+from+mouse+embryonic+and+adult+fibroblast+cultures+by+defined+factors.">Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors.</a> Cell. 126 (4), 663-676 (2006).</li><li>Lian, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Robust+cardiomyocyte+differentiation+from+human+pluripotent+stem+cells+via+temporal+modulation+of+canonical+Wnt+signaling.">Robust cardiomyocyte differentiation from human pluripotent stem cells via temporal modulation of canonical Wnt signaling.</a> Proc Natl Acad Sci. 109 (27), E1848-E1857 (2012).</li><li>Loh, K. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mapping+the+Pairwise+Choices+Leading+from+Pluripotency+to+Human+Bone,+Heart,+and+Other+Mesoderm+Cell+Types.">Mapping the Pairwise Choices Leading from Pluripotency to Human Bone, Heart, and Other Mesoderm Cell Types.</a> Cell. 166 (2), 451-467 (2016).</li><li>Yoshida, Y., Yamanaka, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=iPS+cells:+A+source+of+cardiac+regeneration.">iPS cells: A source of cardiac regeneration.</a> J. Mol. Cell. Cardiol. 50 (2), 327-332 (2011).</li><li>Shiba, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Allogeneic+transplantation+of+iPS+cell-derived+cardiomyocytes+regenerates+primate+hearts.">Allogeneic transplantation of iPS cell-derived cardiomyocytes regenerates primate hearts.</a> Nature. 538, 388-391 (2016).</li><li>Ieda, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+reprogramming+of+fibroblasts+into+functional+cardiomyocytes+by+defined+factors.">Direct reprogramming of fibroblasts into functional cardiomyocytes by defined factors.</a> Cell. 142 (3), 357-386 (2010).</li><li>Song, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heart+repair+by+reprogramming+non-myocytes+with+cardiac+transcription+factors.">Heart repair by reprogramming non-myocytes with cardiac transcription factors.</a> Nature. 485, 599-604 (2012).</li><li>Hirai, H., Katoku-Kikyo, N., Keirstead, S. A., Kikyo, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accelerated+direct+reprogramming+of+fibroblasts+into+cardiomyocyte-like+cells+with+the+MyoD+transactivation+domain.">Accelerated direct reprogramming of fibroblasts into cardiomyocyte-like cells with the MyoD transactivation domain.</a> Cardiovasc Res. 100 (1), 105-113 (2013).</li><li>Qian, L., Berry, E. C., Fu, J., Ieda, M., Srivastava, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reprogramming+of+mouse+fibroblasts+into+cardiomyocyte-like+cells+in+vitro.">Reprogramming of mouse fibroblasts into cardiomyocyte-like cells in vitro.</a> Nat Protoc. 8 (6), 1204-1215 (2013).</li><li>Addis, R. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimization+of+direct+fibroblast+reprogramming+to+cardiomyocytes+using+calcium+activity+as+a+functional+measure+of+success.">Optimization of direct fibroblast reprogramming to cardiomyocytes using calcium activity as a functional measure of success.</a> J Mol Cell Cardiol. 60, 97-106 (2013).</li><li>Ifkovitz, J. L., Addis, R. C., Epstein, J. A., Gearhart, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+TGF&#946;+signaling+increases+direct+conversion+of+fibroblasts+to+induced+cardiomyocytes.">Inhibition of TGF&#946; signaling increases direct conversion of fibroblasts to induced cardiomyocytes.</a> PLoS One. 9 (2), e89678 (2014).</li><li>Zhou, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bmi1+Is+a+Key+Epigenetic+Barrier+to+Direct+Cardiac+Reprogramming.">Bmi1 Is a Key Epigenetic Barrier to Direct Cardiac Reprogramming.</a> Cell Stem Cell. 18 (3), 382-395 (2016).</li><li>Christoforou, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transcription+factors+MYOCD,+SRF,+Mesp1+and+SMARCD3+enhance+the+cardio-inducing+effect+of+GATA4,+TBX5,+and+MEF2C+during+direct+cellular+reprogramming.">Transcription factors MYOCD, SRF, Mesp1 and SMARCD3 enhance the cardio-inducing effect of GATA4, TBX5, and MEF2C during direct cellular reprogramming.</a> PLoS One. 8 (5), e63577 (2013).</li><li>Jayawardena, T. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MicroRNA+induced+cardiac+reprogramming+in+vivo:+evidence+for+mature+cardiac+myocytes+and+improved+cardiac+function.">MicroRNA induced cardiac reprogramming in vivo: evidence for mature cardiac myocytes and improved cardiac function.</a> Circ Res. 116 (3), 418-424 (2015).</li><li>Jayawardena, T. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MicroRNA-mediated+in+vitro+and+in+vivo+direct+reprogramming+of+cardiac+fibroblasts+to+cardiomyocytes.">MicroRNA-mediated in vitro and in vivo direct reprogramming of cardiac fibroblasts to cardiomyocytes.</a> Circ Res. 110 (11), 1465-1473 (2012).</li><li>Cao, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conversion+of+human+fibroblasts+into+functional+cardiomyocytes+by+small+molecules.">Conversion of human fibroblasts into functional cardiomyocytes by small molecules.</a> Science. 352 (6290), 1216-1220 (2016).</li><li>Zhao, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-efficiency+reprogramming+of+fibroblasts+into+cardiomyocytes+requires+suppression+of+pro-fibrotic+signalling.">High-efficiency reprogramming of fibroblasts into cardiomyocytes requires suppression of pro-fibrotic signalling.</a> Nature Communications. 6, 1-15 (2015).</li><li>Morita, S., Kojima, T., Kitamura, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plat-E:+an+efficient+and+stable+system+for+transient+packaging+of+retroviruses.">Plat-E: an efficient and stable system for transient packaging of retroviruses.</a> Gene Therapy. 7, 1063-1066 (2000).</li><li>Zhou, H., Dickson, M. E., Kim, M. S., Bassel-Duby, R., Olson, E. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Akt1/protein+kinase+B+enhances+reprogramming+of+fibroblasts+to+functional+cardiomyocytes.">Akt1/protein kinase B enhances reprogramming of fibroblasts to functional cardiomyocytes.</a> Proc Natl Acad Sci. 112 (38), 11864-11869 (2015).</li><li>Zhou, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ZFN281+enhances+cardiac+reprogramming+by+modulating+cardiac+and+inflammatory+gene+expression.">ZFN281 enhances cardiac reprogramming by modulating cardiac and inflammatory gene expression.</a> Genes &amp; Dev. 31, 1770-1783 (2017).</li><li>Mohamed, T. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+Enhancement+of+In+Vitro+and+In+Vivo+Direct+Cardiac+Reprogramming.">Chemical Enhancement of In Vitro and In Vivo Direct Cardiac Reprogramming.</a> Circulation. 135, 978-995 (2017).</li><li>Qian, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+reprogramming+of+murine+cardiac+fibroblasts+into+induced+cardiomyocytes.">In vivo reprogramming of murine cardiac fibroblasts into induced cardiomyocytes.</a> Nature. 485, 593-598 (2012).</li><li>Mathison, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+situ+reprogramming+to+transdifferentiate+fibroblasts+into+cardiomyocytes+using+adenoviral+vectors:+Implications+for+clinical+myocardial+regeneration.">In situ reprogramming to transdifferentiate fibroblasts into cardiomyocytes using adenoviral vectors: Implications for clinical myocardial regeneration.</a> J Thorac Cardiovasc Surg. , 329-339 (2017).</li></ol>

Föreliggande studie beskriver en högeffektiv strategi för att programmera direkt fibroblaster i funktionella iCMs via leverans av GHMT2m omprogrammering faktorer kombinerat med dämpning av pro-fibrotiska signalvägar. Med hjälp av flödescytometri, Immunofluorescerande imaging, kalcium imaging, och slog blodkroppar, visar vi flertalet celler i detta protokoll genomgå framgångsrika omprogrammering och anta CM härstamning öde. Vi har tidigare visat att tillägg av anti fibrotiska föreningar inklusive TGF-β typ j...

Ischemisk hjärtsjukdom är en ledande dödsorsaken i USA1. Cirka 800 000 amerikaner upplever en första eller återkommande hjärtinfarkt (MI) per år1. Efter MI försämra död hjärtmuskelcellerna (CMs) och hjärtfibros, deponeras av aktiverade hjärt fibroblaster, hjärtat funktion2,3. Progression av hjärtsvikt efter MI är i hög grad oåterkallelig på grund av dålig regenerativ kapacitet av vuxen CMs4,5. Medan nuvarande kliniska behandlingar bromsa sjukdomsförloppet och minska risken för framtida hjärt händelser6,7,8,9, vända inga terapier sjukdomsförloppet på grund av oförmåga att regenerera CMs efter hjärtinfarkt10. Ny cellterapi växer fram för att behandla patienter efter MI. nedslående, kliniska prövningar att leverera stamceller till hjärtat efter MI hittills har visat tveksam regenerativ potentiella11,12, 13 , 14 , 15 , 16 , 17 , 18.
Generering av människa inducerade pluripotenta stamceller (hiPSCs) från fibroblaster av överuttryck av fyra transkriptionsfaktorer, först visat av Takahashi & Yamanaka, öppnade dörren till nya genombrott i cell terapi19. Dessa celler kan differentieras till alla tre groddar lager19, och flera mycket effektiva metoder för att generera ett stort antal CMs har visats tidigare20,21. HiPSC-derived CMs (höfter-CMs) erbjuder en kraftfull plattform för att studera cardiomyogenesis och kan få viktiga konsekvenser för Reparera hjärtat efter skada. Men höfter-CMs står för närvarande inför translationell häck på grund av oro teratoma bildandet22och deras omogna natur kan vara pro-arytmogena23. Omprogrammering fibroblaster i hiPSCs väckte intresse för direkt omprogrammering fibroblaster till andra celltyper. Ieda et al. visade att överuttryck av GATA4, Mef2c och Tbx5 (GMT) i fibroblaster resulterar i direkta omprogrammering till hjärt härstamning, om än i låg effektivitet24. Omprogrammering effektivitet har förbättrats med tillägg av Hand2 (GHMT)25. Sedan dessa tidiga studier, har många publikationer visat att förändra omplanering faktor cocktail med ytterligare transkription faktorer26,27,28,29, kromatin modifierare30,31, mikroRNA32,33eller små molekyler34 leder till förbättrad omprogrammering effektivitet eller mognaden av inducerad hjärtmuskelcellen-liknande celler (iCMs ).
Här tillhandahåller vi ett detaljerat protokoll för att generera iCMs från mus embryonala fibroblaster (MEFs) med hög verkningsgrad. Vi visade tidigare att GHMT cocktail är betydligt bättre med tillägg av miR-1 och miR-133 (GHMT2m) och förbättras ytterligare när pro-fibrotiska signalering vägar, inklusive omvandla tillväxtfaktor β (TGF-β) signalering eller Rho-associerade protein kinase (ROCK) signalvägar är hämmad35. Använder det här protokollet, visar vi att cirka 60% av cellerna express hjärt Troponin T (cTnT), cirka 50% express α-actinin och ett stort antal stryk celler kan observeras så tidigt som dagen 11 efter transduktion av omprogrammering faktorer och behandling med av TGF-β typ I receptor hämmare A-83-01. Dessutom dessa iCMs express gap junction proteiner inklusive connexin 43 och uppvisar spontan kontraktion och kalcium transienter. Detta markerad förbättring i omprogrammering effektivitet jämfört med tidigare studier visar potential att regenerera CMs från endogena cellpopulationer som finns kvar i det efter hjärtinfarkt.

<table border="0" cellpadding="0" cellspacing="0" width="1326">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">C57BL/6 Mice</td>
			<td style="width:197px;">Charles River&#39;s Laboratory</td>
			<td style="width:111px;">027</td>
			<td style="width:667px;">For MEF isolation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Platinum E (PE) Cells</td>
			<td>Cell Biolabs, INC</td>
			<td>RV-101</td>
			<td>For retrovirus production</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">DMEM High Glucose</td>
			<td>Gibco</td>
			<td>SH30022.FS</td>
			<td>Component of iCM, PE, and Growth media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Medium 199</td>
			<td>Life Technologies</td>
			<td>11150-059</td>
			<td>Component of iCM media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fetal Bovine Serum</td>
			<td>Gemini</td>
			<td>100106</td>
			<td>Component of iCM, PE, and Growth media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Donor Horse Serum</td>
			<td>Gemini</td>
			<td>100508 500</td>
			<td>Component of iCM media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">MEM Essential Amino Acids, 50X</td>
			<td>Life Technologies</td>
			<td>11130051</td>
			<td>Component of iCM media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">Sodium Pyruvate Solution, 100X</td>
			<td>Life Technologies</td>
			<td>11360070</td>
			<td>Component of iCM media and for calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">MEM Non-Essential Amino Acids, 100X</td>
			<td>Life Technologies</td>
			<td>11140050</td>
			<td>Component of iCM media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">MEM Vitamin Solution, 100X</td>
			<td>Life Technologies</td>
			<td>11120-052</td>
			<td>Component of iCM media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">Insulin-Transferrin-Selenium</td>
			<td>Gibco</td>
			<td>41400045</td>
			<td>Component of iCM media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">B27</td>
			<td>Gibco</td>
			<td>17504-044</td>
			<td>Component of iCM media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">Penicilin-Streptomycin</td>
			<td>Gibco</td>
			<td>15140-122</td>
			<td>Component of iCM, PE, and Growth media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">GlutaMAX (L-Glutamine Supplement)</td>
			<td>Gibco</td>
			<td>35050-061</td>
			<td>Component of iCM, PE, and Growth media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Blasticidin-HCl</td>
			<td>Life Technologies</td>
			<td>A11139-03</td>
			<td>Component of PE media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">Puromycin dihydrochloride</td>
			<td>Life Technologies</td>
			<td>A11138-03</td>
			<td>Component of PE media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">0.25% Trypsin/EDTA</td>
			<td>Gibco</td>
			<td>25200-056</td>
			<td>For detaching cells from culture dishes</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">A-83-01</td>
			<td>R&#38;D Systems - Tocris</td>
			<td>2939/10</td>
			<td>Treat cells to inhibit TGF-&#946; signaling - promotes high efficiecy reprogramming. Use at 0.5 &#181;M</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">DMSO</td>
			<td>Thermo Scientific</td>
			<td>85190</td>
			<td>For dilution and storage of A-83-01 and component of Freeze Medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">SureCoat</td>
			<td>Cellutron</td>
			<td>SC-9035</td>
			<td>For coating dishes to plate MEFs</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">FuGENE 6 Transfection Reagent</td>
			<td>Promega</td>
			<td>E2692</td>
			<td>Transfection Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Opti-MEM Reduced Serum Media</td>
			<td>Gibco</td>
			<td>11058-021</td>
			<td>Transfection Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pBabe-X Myc-GATA4</td>
			<td></td>
			<td></td>
			<td>Plasmid containing reprogramming factor</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pBabe-X Myc-Hand2</td>
			<td></td>
			<td></td>
			<td>Plasmid containing reprogramming factor</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pBabe-X Myc-Mef2c</td>
			<td></td>
			<td></td>
			<td>Plasmid containing reprogramming factor</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pBabe-X Myc-Tbx5</td>
			<td></td>
			<td></td>
			<td>Plasmid containing reprogramming factor</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pBabe-X miR-1</td>
			<td></td>
			<td></td>
			<td>Plasmid containing reprogramming factor</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pBabe-X miR-133</td>
			<td></td>
			<td></td>
			<td>Plasmid containing reprogramming factor</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">pBabe-X GFP</td>
			<td></td>
			<td></td>
			<td>Plasmid containing reprogramming factor</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Polybrene (Hexadimethrine bromide)</td>
			<td>Sigma</td>
			<td>H9268-5G</td>
			<td>For viral induction. Use at a concentration of 6 &#181;g/mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">Vacuum Filter + bottles (0.22 &#181;m pores)</td>
			<td>Nalgene</td>
			<td>&#160;569-0020&#160;</td>
			<td>For filtering media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Syringes</td>
			<td>Bd Vacutainer Labware&#160;</td>
			<td>309654</td>
			<td>For viral filtration</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">0.45 &#181;m Filters</td>
			<td>Celltreat</td>
			<td>229749</td>
			<td>For viral filtration</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">70 &#181;m cell strainers</td>
			<td>Falcon&#160;</td>
			<td>352350</td>
			<td>For MEF isolation and Flow Cytometry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">Cytofix/Cytoperm Solution</td>
			<td>BD</td>
			<td>554722</td>
			<td>For fixation and permeabilization of cells for flow cytometry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">perm/wash buffer&#160;</td>
			<td>BD</td>
			<td>554723</td>
			<td>For washing cells for flow cytometry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">DPBS 1X</td>
			<td>Gibco</td>
			<td>14190-250</td>
			<td>For washing cells</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bovine Serum Albumin</td>
			<td>VWR</td>
			<td>0332-100g</td>
			<td>For flow cytometry and calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Goat Serum</td>
			<td>Sigma&#160;</td>
			<td>G9023</td>
			<td>For blocking cells for Flow Cytometry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Donkey Serum</td>
			<td>Sigma</td>
			<td>D9663-10mg</td>
			<td>For blocking cells for Flow Cytometry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mouse Troponin T</td>
			<td>Thermo Scientific</td>
			<td>ms-295-p</td>
			<td>1:400 IF, 1:200 Flow Cytometry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mouse &#945;-actinin</td>
			<td>Sigma</td>
			<td>A7811L</td>
			<td>1:400 IF, 1:200 Flow Cytometry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rabbit Connexin 43</td>
			<td>Sigma&#160;</td>
			<td>C6219</td>
			<td>1:400 IF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rabbit Troponin I</td>
			<td>PhosphoSolutions</td>
			<td>2010-TNI</td>
			<td>1:400 IF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hoechst</td>
			<td>Life Technologies</td>
			<td>62249</td>
			<td>1:10000 IF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Alexa 488, rabbit</td>
			<td>Life Technologies</td>
			<td>A-11034</td>
			<td>1:800 IF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Alexa 555, mouse</td>
			<td>Life Technologies</td>
			<td>A-21422</td>
			<td>1:800 IF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Alexa 647, mouse</td>
			<td>Life Technologies</td>
			<td>A-31571</td>
			<td>1:200 Flow Cytometry</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">27-color ZE5 Flow Cytometer&#160;</td>
			<td>Bio-RAD</td>
			<td></td>
			<td>For FACS</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Paraformaldehyde</td>
			<td>sigma</td>
			<td>P6148-500mg</td>
			<td>For fixing cells for IF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Triton X-100</td>
			<td>Promega</td>
			<td>H5142</td>
			<td>For permeabilization of cells for IF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">EVOS&#8482; FL Color Imaging System</td>
			<td>Thermo Scientific</td>
			<td>AMEFC4300</td>
			<td>For IF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">NaCl</td>
			<td>RPI</td>
			<td>S23020-5000</td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">KCl</td>
			<td>VWR</td>
			<td>395</td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">CaCl2</td>
			<td>Fisher</td>
			<td>C614-500</td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">MgCl2</td>
			<td>VWR</td>
			<td>97061-352</td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">glucose</td>
			<td>sigma</td>
			<td>G7528-250g</td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">HEPES</td>
			<td>sigma</td>
			<td>H4034-500g</td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fura-2 AM</td>
			<td>Life Technologies</td>
			<td>F1221</td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fluronic F-127</td>
			<td>Sigma</td>
			<td>P2443-250g</td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Nifedipine</td>
			<td>Sigma</td>
			<td>N7634-1G</td>
			<td>For disruption calcium transients in iCMs - use at 10 &#181;M</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoproterenol</td>
			<td>sigma</td>
			<td>I6504-1g</td>
			<td>For increasing number of calcium transients in iCMs - use at 1-2 &#181;M</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Marianas Spinning Disk Confocal microscope</td>
			<td>3i</td>
			<td></td>
			<td>For calcium imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ethanol</td>
			<td>Decon Laboratories</td>
			<td>2801</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">bleach</td>
			<td>Clorox</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">50 mL conical tubes</td>
			<td>GREINER BIO-ONE</td>
			<td>227261</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">15 mL conical tubes</td>
			<td>GREINER BIO-ONE</td>
			<td>188271</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">15 cm cell culture dishes</td>
			<td>Falcon</td>
			<td>353025</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10 cm cell culture dishes</td>
			<td>Falcon</td>
			<td>353003</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">60 mm cell culture dishes</td>
			<td>GREINER BIO-ONE</td>
			<td>628160</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">6 well cell culture plates</td>
			<td>GREINER BIO-ONE</td>
			<td>657160&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">12 well cell culture plates</td>
			<td>GREINER BIO-ONE</td>
			<td>665180&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">24 well cell culture plates</td>
			<td>GREINER BIO-ONE</td>
			<td>662160&#160;</td>
			<td></td>
		</tr>
	</tbody>
</table>

suppression of pro-fibrotic signaling potentiates factor-mediated reprogramming of mouse embryonic fibroblasts into induced cardiomyocytes

Trans-differentiering av en somatisk celltyp till en annan har en enorm potential att modellera och behandla sjukdomar. Tidigare studier har visat att mus embryonala, dermal, och hjärtriskfaktorer fibroblaster kan omprogrammeras till funktionella inducerad-hjärtmuskelcellen-liknande celler (iCMs) genom överuttryck av kardiogen transkriptionsfaktorer inklusive GATA4, Hand2, Mef2c, och Tbx5 både in vitro- och in vivo. Dessa tidigare studier har dock visat relativt låg effektivitet. För att återställa hjärtats funktion efter skada, måste mekanismer som styr hjärtats omprogrammering belysas för att öka effektiviteten och mognaden av iCMs.
Vi har tidigare visat att hämning av pro-fibrotiska signalering dramatiskt ökar omplanering effektivitet. Här, detalj vi metoder för att uppnå en omplanering verkningsgrad på upp till 60%. Dessutom beskriver vi flera metoder inklusive flödescytometri, Immunofluorescerande imaging och kalcium imaging att kvantifiera omplanering effektivitet och mognaden av omprogrammerade fibroblaster. Med hjälp av protokollet som beskrivs här, kan mekanistiska studier genomföras att fastställa positiva och negativa regulatorer av hjärt omprogrammering. Dessa studier kan identifiera signalvägar som kan vara inriktade på att främja omplanering effektivitet och mognad, vilket kan leda till nya cell terapier att behandla mänskliga hjärtsjukdom.

Använder den omprogrammering strategi som beskrivs ovan och i figur 1B, genererade vi iCMs med cirka 70% av celler som uttrycker hjärt Troponin T och cirka 55% av celler som uttrycker hjärt α-actinin, kvantifieras av flödescytometri på dag 9 efter transduktion av GHMT2m (figur 2A och B). Dessutom flertalet celler express hjärt Troponin T, Troponin I, och hjärt α-actinin samt gap junction markören Connex...

Watch this Scientific Journal Video about Suppression of Pro-fibrotic Signaling Potentiates Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts into Induced Cardiomyocytes at JoVE.com

Suppression of Pro-fibrotic Signaling Potentiates Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts into Induced Cardiomyocytes

Här presenterar vi en robust metod för att programmera primära embryonala fibroblaster i funktionella hjärtmuskelceller genom överuttryck av GATA4, Hand2, Mef2c, Tbx5, miR-1 och miR-133 (GHMT2m) tillsammans med hämning av TGF-β signalering. Våra protokoll genererar slående hjärtmuskelceller så tidigt som 7 dagar efter transduktion med upp till 60% effektivitet.

Dämpning av Pro-fibrotiska signalering potentierar faktor-medierad omprogrammering av mus embryonala fibroblaster till inducerad hjärtmuskelcellerna

Trans-differentiation of one somatic cell type into another has enormous potential to model and treat human diseases. Previous studies have shown that ...

Research

JoVE Journal

Developmental Biology

Stable and Efficient Genetic Modification of Cells in the Adult Mouse V-SVZ for the Analysis of Neural Stem Cell Autonomous and Non-autonomous Effects

Stabil och effektiv genetisk modifiering av celler i vuxen mus V-SVZ för analys av neurala stamceller autonoma och icke-självständiga effekter

Relatively quiescent somatic stem cells support life-long cell renewal in most adult tissues. Neural stem cells in the adult mammalian brain are ...

Improved Generation of Induced Cardiomyocytes Using a Polycistronic Construct Expressing Optimal Ratio of Gata4, Mef2c and Tbx5

Förbättrad Generering av inducerade Cardiomyocytes Använda en polycistroniskt konstruktion som uttrycker optimala förhållandet mellan GATA4, Mef2c och Tbx5

Direct conversion of cardiac fibroblasts (CFs) into induced cardiomyocytes (iCMs) holds great potential for regenerative medicine by offering alternative ...

Protocol for the Direct Conversion of Murine Embryonic Fibroblasts into Trophoblast Stem Cells

Protokoll för direkt omvandling av Murina embryonala fibroblaster i trofoblaster stamceller

Trophoblast stem cells (TSCs) arise as a consequence of the first cell fate decision in mammalian development. They can be cultured in vitro, retaining ...

Reprogramming Mouse Embryonic Fibroblasts with Transcription Factors to Induce a Hemogenic Program

Omprogrammering Mouse Embryonala fibroblasts med transkriptionsfaktorer att inducera en Hemogenic Program

This protocol details the induction of a hemogenic program in mouse embryonic fibroblasts (MEFs) via overexpression of transcription factors (TFs). We ...

Stimulation of Notch Signaling in Mouse Osteoclast Precursors

Stimulering av Notch signalering i mus osteoklastprekursorer

Notch signaling is a key component of multiple physiological and pathological processes. The nature of Notch signaling, however, makes in vitro ...

Assessing Cardiomyocyte Subtypes Following Transcription Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts

Uppskatta cardiomyocyte Subtyper Efter transkriptionsfaktor-medierad Omprogrammering av mus embryonala fibroblaster

Direct reprogramming of one cell type into another has recently emerged as a powerful paradigm for regenerative medicine, disease modeling, and lineage ...

Analysis of Retinoic Acid-induced Neural Differentiation of Mouse Embryonic Stem Cells in Two and Three-dimensional Embryoid Bodies

Analys av retinsyra-inducerade Neural differentiering av Mouse embryonala stamceller i två och tre-dimensionella Embryoid organ

Mouse embryonic stem cells (ESCs) isolated from the inner mass of the blastocyst (typically at day E3.5), can be used as in vitro model system for ...

Dissection and Culture of Mouse Embryonic Kidney

Dissektion och kultur av mus embryonisk njure

The goal of this protocol is to describe a method for the dissection, isolation, and culture of mouse metanephric rudiments. 
During mammalian kidney ...

Evaluation of Injury-induced Senescence and In Vivo Reprogramming in the Skeletal Muscle

Evaluation of Injury-induced Senescence and In Vivo Reprogramming in the Skeletal Muscle

Utvärdering av nervskade-inducerad åldras och In Vivo omprogrammering i skelettmuskulaturen

Cellular senescence is a stress response that is characterized by a stable cellular growth arrest, which is important for many physiological and ...

Direct Lineage Reprogramming of Adult Mouse Fibroblast to Erythroid Progenitors

Direkt härstamning omprogrammering av vuxen mus Fibroblast till erytroid stamfäder

Erythroid cell commitment and differentiation proceed through activation of a lineage-restricted transcriptional network orchestrated by a group of cell ...