A simplified, Langendorff-free method for concomitant isolation of viable cardiac myocytes and nonmyocytes from the adult mouse heart

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A simplified, Langendorff-free method for concomitant isolation of viable cardiac myocytes and nonmyocytes from the adult mouse heart. / Ackers-Johnson, M; Li, Peter Yiqing; Holmes, Andrew; O'Brien, Sian-Marie; Pavlovic, Davor; Foo, Roger.

In: Circulation Research, Vol. 119, No. 8, 30.09.2016, p. 909-920.

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@article{dd51691b1ceb40ae9824d09b49197191,
title = "A simplified, Langendorff-free method for concomitant isolation of viable cardiac myocytes and nonmyocytes from the adult mouse heart",
abstract = "Rationale: 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 specialised Langendorff apparatus, which poses considerable logistical and technical barriers to researchers, and demands extensive training investment.Objective: To identify and optimise a convenient, alternative approach, allowing the robust isolation and culture of adult mouse cardiac myocytes using only common surgical and laboratory equipment.Methods and Results: 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 organised contractile and mitochondrial morphology, transcriptional signatures, calcium handling, responses to hypoxia, neurohormonal stimulation and electrical pacing, and are amenable to patch clamp and adenoviral gene transfer techniques. Furthermore, the methodology permits concurrent isolation, separation and co-culture of myocyte and non-myocyte cardiac populations.Conclusions: We present a novel, simplified method, demonstrating concomitant isolation of viable cardiac myocytes and non-myocytes from the same adult mouse heart. We anticipate that this new approach will expand and accelerate innovative research in the field of cardiac biology.",
keywords = "Langendorff, cell isolation, cardiac myocyte, cell culture, fibroblasts",
author = "M Ackers-Johnson and Li, {Peter Yiqing} and Andrew Holmes and Sian-Marie O'Brien and Davor Pavlovic and Roger Foo",
year = "2016",
month = sep,
day = "30",
doi = "10.1161/CIRCRESAHA.116.309202",
language = "English",
volume = "119",
pages = "909--920",
journal = "Circulation Research",
issn = "0009-7330",
publisher = "American Heart Association",
number = "8",

}

RIS

TY - JOUR

T1 - A simplified, Langendorff-free method for concomitant isolation of viable cardiac myocytes and nonmyocytes from the adult mouse heart

AU - Ackers-Johnson, M

AU - Li, Peter Yiqing

AU - Holmes, Andrew

AU - O'Brien, Sian-Marie

AU - Pavlovic, Davor

AU - Foo, Roger

PY - 2016/9/30

Y1 - 2016/9/30

N2 - Rationale: 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 specialised Langendorff apparatus, which poses considerable logistical and technical barriers to researchers, and demands extensive training investment.Objective: To identify and optimise a convenient, alternative approach, allowing the robust isolation and culture of adult mouse cardiac myocytes using only common surgical and laboratory equipment.Methods and Results: 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 organised contractile and mitochondrial morphology, transcriptional signatures, calcium handling, responses to hypoxia, neurohormonal stimulation and electrical pacing, and are amenable to patch clamp and adenoviral gene transfer techniques. Furthermore, the methodology permits concurrent isolation, separation and co-culture of myocyte and non-myocyte cardiac populations.Conclusions: We present a novel, simplified method, demonstrating concomitant isolation of viable cardiac myocytes and non-myocytes from the same adult mouse heart. We anticipate that this new approach will expand and accelerate innovative research in the field of cardiac biology.

AB - Rationale: 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 specialised Langendorff apparatus, which poses considerable logistical and technical barriers to researchers, and demands extensive training investment.Objective: To identify and optimise a convenient, alternative approach, allowing the robust isolation and culture of adult mouse cardiac myocytes using only common surgical and laboratory equipment.Methods and Results: 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 organised contractile and mitochondrial morphology, transcriptional signatures, calcium handling, responses to hypoxia, neurohormonal stimulation and electrical pacing, and are amenable to patch clamp and adenoviral gene transfer techniques. Furthermore, the methodology permits concurrent isolation, separation and co-culture of myocyte and non-myocyte cardiac populations.Conclusions: We present a novel, simplified method, demonstrating concomitant isolation of viable cardiac myocytes and non-myocytes from the same adult mouse heart. We anticipate that this new approach will expand and accelerate innovative research in the field of cardiac biology.

KW - Langendorff

KW - cell isolation

KW - cardiac myocyte

KW - cell culture

KW - fibroblasts

U2 - 10.1161/CIRCRESAHA.116.309202

DO - 10.1161/CIRCRESAHA.116.309202

M3 - Article

VL - 119

SP - 909

EP - 920

JO - Circulation Research

JF - Circulation Research

SN - 0009-7330

IS - 8

ER -