{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,27]],"date-time":"2026-10-27T22:00:53Z","timestamp":1793138453472,"version":"4.5.0"},"reference-count":68,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"8","content-domain":{"domain":["www.ahajournals.org"],"crossmark-restriction":true},"short-container-title":["Circulation Research"],"published-print":{"date-parts":[[2016,9,30]]},"abstract":"<jats:sec>\n                    <jats:title>\n                      <jats:underline>Rationale:<\/jats:underline>\n                    <\/jats:title>\n                    <jats:p>Cardiovascular disease represents a global pandemic. The advent of and recent advances in mouse genomics, epigenomics, and transgenics offer ever-greater potential for powerful avenues of research. However, progress is often constrained by unique complexities associated with the isolation of viable myocytes from the adult mouse heart. Current protocols rely on retrograde aortic perfusion using specialized Langendorff apparatus, which poses considerable logistical and technical barriers to researchers and demands extensive training investment.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>\n                      <jats:underline>Objective:<\/jats:underline>\n                    <\/jats:title>\n                    <jats:p>To identify and optimize a convenient, alternative approach, allowing the robust isolation and culture of adult mouse cardiac myocytes using only common surgical and laboratory equipment.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>\n                      <jats:underline>Methods and Results:<\/jats:underline>\n                    <\/jats:title>\n                    <jats:p>Cardiac myocytes were isolated with yields comparable to those in published Langendorff-based methods, using direct needle perfusion of the LV ex vivo and without requirement for heparin injection. Isolated myocytes can be cultured antibiotic free, with retained organized contractile and mitochondrial morphology, transcriptional signatures, calcium handling, responses to hypoxia, neurohormonal stimulation, and electric pacing, and are amenable to patch clamp and adenoviral gene transfer techniques. Furthermore, the methodology permits concurrent isolation, separation, and coculture of myocyte and nonmyocyte cardiac populations.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>\n                      <jats:underline>Conclusions:<\/jats:underline>\n                    <\/jats:title>\n                    <jats:p>We present a novel, simplified method, demonstrating concomitant isolation of viable cardiac myocytes and nonmyocytes from the same adult mouse heart. We anticipate that this new approach will expand and accelerate innovative research in the field of cardiac biology.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1161\/circresaha.116.309202","type":"journal-article","created":{"date-parts":[[2016,8,8]],"date-time":"2016-08-08T23:03:38Z","timestamp":1470697418000},"page":"909-920","update-policy":"https:\/\/doi.org\/10.1161\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":572,"title":["A Simplified, Langendorff-Free Method for Concomitant Isolation of Viable Cardiac Myocytes and Nonmyocytes From the Adult Mouse Heart"],"prefix":"10.1161","volume":"119","author":[{"given":"Matthew","family":"Ackers-Johnson","sequence":"first","affiliation":[{"name":"From the Cardiovascular Research Institute, Centre for Translational Medicine MD6, National University Health System, Singapore (M.A.-J., P.Y.L., R.S.F.); Genome Institute of Singapore (M.A.-J., R.S.F.); and Institute of Cardiovascular Sciences, University of Birmingham, UK (A.P.H., S.-M.O., D.P.)."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter Yiqing","family":"Li","sequence":"additional","affiliation":[{"name":"From the Cardiovascular Research Institute, Centre for Translational Medicine MD6, National University Health System, Singapore (M.A.-J., P.Y.L., R.S.F.); Genome Institute of Singapore (M.A.-J., R.S.F.); and Institute of Cardiovascular Sciences, University of Birmingham, UK (A.P.H., S.-M.O., D.P.)."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew P.","family":"Holmes","sequence":"additional","affiliation":[{"name":"From the Cardiovascular Research Institute, Centre for Translational Medicine MD6, National University Health System, Singapore (M.A.-J., P.Y.L., R.S.F.); Genome Institute of Singapore (M.A.-J., R.S.F.); and Institute of Cardiovascular Sciences, University of Birmingham, UK (A.P.H., S.-M.O., D.P.)."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sian-Marie","family":"O\u2019Brien","sequence":"additional","affiliation":[{"name":"From the Cardiovascular Research Institute, Centre for Translational Medicine MD6, National University Health System, Singapore (M.A.-J., P.Y.L., R.S.F.); Genome Institute of Singapore (M.A.-J., R.S.F.); and Institute of Cardiovascular Sciences, University of Birmingham, UK (A.P.H., S.-M.O., D.P.)."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Davor","family":"Pavlovic","sequence":"additional","affiliation":[{"name":"From the Cardiovascular Research Institute, Centre for Translational Medicine MD6, National University Health System, Singapore (M.A.-J., P.Y.L., R.S.F.); Genome Institute of Singapore (M.A.-J., R.S.F.); and Institute of Cardiovascular Sciences, University of Birmingham, UK (A.P.H., S.-M.O., D.P.)."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roger S.","family":"Foo","sequence":"additional","affiliation":[{"name":"From the Cardiovascular Research Institute, Centre for Translational Medicine MD6, National University Health System, Singapore (M.A.-J., P.Y.L., R.S.F.); Genome Institute of Singapore (M.A.-J., R.S.F.); and Institute of Cardiovascular Sciences, University of Birmingham, UK (A.P.H., S.-M.O., D.P.)."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"276","reference":[{"key":"e_1_3_5_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrcardio.2014.26"},{"key":"e_1_3_5_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijcard.2013.12.028"},{"key":"e_1_3_5_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(14)61889-4"},{"key":"e_1_3_5_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature01306"},{"key":"e_1_3_5_6_2","doi-asserted-by":"publisher","DOI":"10.1155\/2015\/482171"},{"key":"e_1_3_5_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/0005-2744(69)90410-0"},{"key":"e_1_3_5_8_2","doi-asserted-by":"publisher","DOI":"10.1161\/01.RES.26.6.679"},{"key":"e_1_3_5_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0008-6363(98)00128-X"},{"key":"e_1_3_5_10_2","doi-asserted-by":"publisher","DOI":"10.1152\/ajpheart.2000.279.1.H429"},{"key":"e_1_3_5_11_2","unstructured":"Alliance for Cellular Signalling (AfCS) Procedure Protocol ID PP00000125. http:\/\/www.signaling-gateway.org\/data\/ProtocolLinks.html."},{"key":"e_1_3_5_12_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-59745-571-8_16"},{"key":"e_1_3_5_13_2","doi-asserted-by":"publisher","DOI":"10.1385\/1-59745-214-9:271"},{"key":"e_1_3_5_14_2","first-page":"e1433","article-title":"Isolation and genetic manipulation of adult cardiac myocytes for confocal imaging.","volume":"31","author":"Kaestner L","year":"2009","unstructured":"Kaestner L, Scholz A, Hammer K, Vecerdea A, Ruppenthal S, Lipp P. Isolation and genetic manipulation of adult cardiac myocytes for confocal imaging. J Vis Exp 2009;31:e1433.","journal-title":"J Vis Exp"},{"key":"e_1_3_5_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.yjmcc.2011.06.012"},{"key":"e_1_3_5_16_2","first-page":"e50289","article-title":"Isolation, culture, and functional characterization of adult mouse cardiomyoctyes.","volume":"79","author":"Graham EL","year":"2013","unstructured":"Graham EL, Balla C, Franchino H, Melman Y, del Monte F, Das S. Isolation, culture, and functional characterization of adult mouse cardiomyoctyes. J Vis Exp 2013;79:e50289.","journal-title":"J Vis Exp"},{"key":"e_1_3_5_17_2","first-page":"e51109","article-title":"Isolation and physiological analysis of mouse cardiomyocytes.","volume":"91","author":"Roth GM","year":"2014","unstructured":"Roth GM, Bader DM, Pfaltzgraff ER. Isolation and physiological analysis of mouse cardiomyocytes. J Vis Exp 2014;91:e51109.","journal-title":"J Vis Exp"},{"key":"e_1_3_5_18_2","first-page":"e51357","article-title":"Isolation and culture of adult mouse cardiomyocytes for cell signaling and in vitro cardiac hypertrophy.","volume":"87","author":"Li D","year":"2014","unstructured":"Li D, Wu J, Bai Y, Zhao X, Liu L. Isolation and culture of adult mouse cardiomyocytes for cell signaling and in vitro cardiac hypertrophy. J Vis Exp 2014;87:e51357.","journal-title":"J Vis Exp"},{"key":"e_1_3_5_19_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-2277.2000.tb01055.x"},{"key":"e_1_3_5_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.40.4.1567-1568.2002"},{"key":"e_1_3_5_21_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00335-012-9411-5"},{"key":"e_1_3_5_22_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature13992"},{"key":"e_1_3_5_23_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.3155"},{"key":"e_1_3_5_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2006.03.014"},{"key":"e_1_3_5_25_2","doi-asserted-by":"publisher","DOI":"10.1152\/physiolgenomics.00129.2012"},{"key":"e_1_3_5_26_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ceca.2007.04.001"},{"key":"e_1_3_5_27_2","first-page":"H238","article-title":"Effect of taurine on calcium paradox and ischemic heart failure.","volume":"240","author":"Kramer JH","year":"1981","unstructured":"Kramer JH, Chovan JP, Schaffer SW. Effect of taurine on calcium paradox and ischemic heart failure. Am J Physiol. 1981;240:H238\u2013H246.","journal-title":"Am J Physiol"},{"key":"e_1_3_5_28_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2828(82)90171-7"},{"key":"e_1_3_5_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2828(91)90103-S"},{"key":"e_1_3_5_30_2","doi-asserted-by":"publisher","DOI":"10.1007\/s12013-011-9165-9"},{"key":"e_1_3_5_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ceca.2005.07.006"},{"key":"e_1_3_5_32_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.yjmcc.2010.05.010"},{"key":"e_1_3_5_33_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF01131017"},{"key":"e_1_3_5_34_2","doi-asserted-by":"publisher","DOI":"10.1161\/01.CIR.0000129305.25115.80"},{"key":"e_1_3_5_35_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.yjmcc.2015.03.007"},{"key":"e_1_3_5_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.devcel.2015.04.021"},{"key":"e_1_3_5_37_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2828(90)90976-9"},{"key":"e_1_3_5_38_2","doi-asserted-by":"publisher","DOI":"10.1152\/ajpheart.00704.2013"},{"key":"e_1_3_5_39_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCULATIONAHA.107.754051"},{"key":"e_1_3_5_40_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.hrthm.2015.11.022"},{"key":"e_1_3_5_41_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.109.194423"},{"key":"e_1_3_5_42_2","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.33.1.103"},{"key":"e_1_3_5_43_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M113.533851"},{"key":"e_1_3_5_44_2","doi-asserted-by":"publisher","DOI":"10.1152\/ajpheart.00068.2015"},{"key":"e_1_3_5_45_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(01)00404-4"},{"key":"e_1_3_5_46_2","doi-asserted-by":"publisher","DOI":"10.1161\/01.RES.0000218781.23144.3e"},{"key":"e_1_3_5_47_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.yjmcc.2013.10.006"},{"key":"e_1_3_5_48_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.117.305349"},{"key":"e_1_3_5_49_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2014.113"},{"key":"e_1_3_5_50_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.115.303794"},{"key":"e_1_3_5_51_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.114.302530"},{"key":"e_1_3_5_52_2","doi-asserted-by":"publisher","DOI":"10.1023\/A:1006802719136"},{"key":"e_1_3_5_53_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-1096-0_19"},{"key":"e_1_3_5_54_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.114.302437"},{"key":"e_1_3_5_55_2","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms8229"},{"key":"e_1_3_5_56_2","first-page":"e3302","article-title":"Single cell transcriptional profiling of adult mouse cardiomyocytes.","volume":"58","author":"Flynn JM","year":"2011","unstructured":"Flynn JM, Santana LF, Melov S. Single cell transcriptional profiling of adult mouse cardiomyocytes. J Vis Exp 2011;58:e3302.","journal-title":"J Vis Exp"},{"key":"e_1_3_5_57_2","doi-asserted-by":"publisher","DOI":"10.1093\/cvr\/cvu112"},{"key":"e_1_3_5_58_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2828(78)90354-1"},{"key":"e_1_3_5_59_2","doi-asserted-by":"publisher","DOI":"10.1007\/s10517-005-0492-y"},{"key":"e_1_3_5_60_2","doi-asserted-by":"publisher","DOI":"10.1161\/01.RES.59.2.143"},{"key":"e_1_3_5_61_2","first-page":"451","article-title":"Selective use of calcium chelators enhances the yield of calcium-tolerant myocytes from adult heart.","volume":"35","author":"Nair P","year":"1997","unstructured":"Nair P, Nair RR. Selective use of calcium chelators enhances the yield of calcium-tolerant myocytes from adult heart. Indian J Exp Biol. 1997;35:451\u2013456.","journal-title":"Indian J Exp Biol"},{"key":"e_1_3_5_62_2","doi-asserted-by":"publisher","DOI":"10.1152\/ajplegacy.1967.213.5.1143"},{"key":"e_1_3_5_63_2","doi-asserted-by":"publisher","DOI":"10.1093\/cvr\/28.8.1193"},{"key":"e_1_3_5_64_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.yjmcc.2013.09.004"},{"key":"e_1_3_5_65_2","doi-asserted-by":"publisher","DOI":"10.1152\/ajpheart.01144.2007"},{"key":"e_1_3_5_66_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.yjmcc.2011.09.001"},{"key":"e_1_3_5_67_2","doi-asserted-by":"publisher","DOI":"10.1152\/ajpheart.1998.275.4.H1260"},{"key":"e_1_3_5_68_2","doi-asserted-by":"publisher","DOI":"10.1023\/A:1022047102064"},{"key":"e_1_3_5_69_2","doi-asserted-by":"publisher","DOI":"10.1161\/01.RES.69.5.1280"}],"container-title":["Circulation Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.ahajournals.org\/doi\/full\/10.1161\/CIRCRESAHA.116.309202","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,5,12]],"date-time":"2024-05-12T23:32:38Z","timestamp":1715556758000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.ahajournals.org\/doi\/10.1161\/CIRCRESAHA.116.309202"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,9,30]]},"references-count":68,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2016,9,30]]}},"alternative-id":["10.1161\/CIRCRESAHA.116.309202"],"URL":"https:\/\/doi.org\/10.1161\/circresaha.116.309202","relation":{},"ISSN":["0009-7330","1524-4571"],"issn-type":[{"value":"0009-7330","type":"print"},{"value":"1524-4571","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,9,30]]},"assertion":[{"value":"2016-05-29","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2016-08-05","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2016-08-08","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}