A Simplified, Langendorff-Free Method for Concomitant Isolation of Viable Cardiac Myocytes and Nonmyocytes From the Adult Mouse Heart. Ackers-Johnson M, Li PY, Holmes AP, O’Brien SM, Pavlovic D, Foo RS. Circ Res 2016.
See also:
Langendorff-Free Isolation and Propagation of Adult Mouse Cardiomyocytes. Ackers-Johnson M, Foo RS. Methods Mol Biol. 2019;1940:193-204.
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Here, we outline a convenient, alternative approach to isolation and culture of viable myocytes and non-myocytes from the adult mouse heart, developed in our lab. The technique requires only common surgical and laboratory equipment, and involves direct needle perfusion of the left ventricle ex vivo. Viable myocyte yields and functionality are comparable to those described in established Langendorff-based protocols.
Since publication, this procedure has been reproduced and established in other labs, worldwide. We continue to collaborate and advise on implementation and troubleshooting, please get in touch!
[trx_image url=”http://www.foo-lab.com/wp-content/uploads/2019/06/Image-2-1024×374.png” title=”Freshly isolated myocytes from an adult mouse heart. Scale bars, 100 µm.” shape=”square” top=”inherit” bottom=”inherit” left=”inherit” right=”inherit”]
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[trx_columns fluid=”no” margins=”yes” top=”inherit” bottom=”inherit” left=”inherit” right=”inherit” count=”2″] [trx_column_item align=”center”][trx_image url=”http://www.foo-lab.com/wp-content/uploads/2019/06/Image-3.png” title=”Progressive remodelling of adult mouse myocytes over 3 weeks in culture. Scale bars, 100 µm.” shape=”square” top=”inherit” bottom=”inherit” left=”inherit” right=”inherit”]
[/trx_column_item] [trx_column_item align=”center”][trx_image url=”http://www.foo-lab.com/wp-content/uploads/2019/06/image-4.jpeg” title=”Co-culture of cardiac myocytes (green) and fibroblasts (red) isolated from a single heart. Scale bar, 100 µm.” shape=”square” top=”Large” bottom=”inherit” left=”inherit” right=”inherit”][/trx_column_item]
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[trx_title type=”4″ style=”divider” align=”center” top=”inherit” bottom=”inherit” left=”inherit” right=”inherit”]Citations[/trx_title]
(Selected citations from 57 to-date)
Glycogen Synthase Kinase-3α Promotes Fatty Acid Uptake and Lipotoxic Cardiomyopathy. Nakamura M, Liu T, Husain S, Zhai P, Warren JS, Hsu CP, Matsuda T, Phiel CJ, Cox JE, Tian B, Li H, Sadoshima J Cell Metab. 2019;29:1119-1134.e12
Robust CTCF-Based Chromatin Architecture Underpins Epigenetic Changes in the Heart Failure Stress-Gene Response. Lee DP, Tan WLW, Anene-Nzelu CG, Lee CJM, Li PY, Luu TDA, Chan CX, Tiang Z, Ng SL, Huang X, Efthymios M, Autio MI, Jiang J, Fullwood MJ, Prabhakar S, Lieberman Aiden E, Foo RS. Circulation. 2019;139:1937-1956
The N6-Methyladenosine mRNA Methylase METTL3 Controls Cardiac Homeostasis and Hypertrophy. Dorn LE, Lasman L, Chen J, Xu X, Hund TJ, Medvedovic M, Hanna JH, van Berlo JH, Accornero F. Circulation. 2019;139:533-545
Megakaryocytic Leukemia 1 Bridges Epigenetic Activation of NADPH Oxidase in Macrophages to Cardiac Ischemia-Reperfusion Injury. Yu L, Yang G, Zhang X, Wang P, Weng X, Yang Y, Li Z, Fang M, Xu Y, Sun A, Ge J. Circulation. 2018;138:2820-2836
Uncoupling protein 3 deficiency impairs myocardial fatty acid oxidation and contractile recovery following ischemia/reperfusion. Edwards KS, Ashraf S, Lomax TM, Wiseman JM, Hall ME, Gava FN, Hall JE, Hosler JP, Harmancey R. Basic Res Cardiol. 2018;113:47
Paracrine effect of regulatory T cells promotes cardiomyocyte proliferation during pregnancy and after myocardial infarction. Zacchigna S, Martinelli V, Moimas S, Colliva A, Anzini M, Nordio A, Costa A, Rehman M, Vodret S, Pierro C, Colussi G, Zentilin L, Gutierrez MI, Dirkx E, Long C, Sinagra G, Klatzmann D, Giacca M. Nat Commun. 2018;9:2432
With or Without Langendorff: A New Method for Adult Myocyte Isolation to Be Tested With Time. Chen X, O’Connell TD, Xiang YK. Circ Res. 2016;119:888-90
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