Isolation of Atrial Myocytes from Adult Mice

Jansen, Hailey J.; Rose, Robert A.

doi:10.3791/59588

Cited by 7 publications

(6 citation statements)

References 21 publications

(52 reference statements)

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“…The protocol described above for calcium reintroduction into the isolated ventricular myocyte cultures was designed to limit cardiac myocyte death via inappropriate calcium influx via store operated calcium channels. It should be noted that the stepwise calcium reintroduction should not be performed for isolated atrial myocyte cultures, as this will promote cell death during shortand long-term culture 12 . For further precaution, the perfusion and digestion buffers used here include the cardiac muscle contraction inhibitor butanedione monoxime (BDM) to avoid hypercontraction of isolated myocytes, as well as the calcium paradox, both of which impact myocyte viability 26 .…”

Section: Discussionmentioning

confidence: 99%

Simultaneous Isolation and Culture of Atrial Myocytes, Ventricular Myocytes, and Non-Myocytes from an Adult Mouse Heart

Blackwood

Bilal

Azizi

et al. 2020

JoVE

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The isolation and culturing of cardiac myocytes from mice has been essential for furthering the understanding of cardiac physiology and pathophysiology. While isolating myocytes from neonatal mouse hearts is relatively straightforward, myocytes from the adult murine heart are preferred. This is because compared to neonatal cells, adult myocytes more accurately recapitulate cell function as it occurs in the adult heart in vivo. However, it is technically difficult to isolate adult mouse cardiac myocytes in the necessary quantities and viability, which contributes to an experimental impasse.Furthermore, published procedures are specific for the isolation of either atrial or ventricular myocytes at the expense of atrial and ventricular non-myocyte cells.Described here is a detailed method for isolating both atrial and ventricular cardiac myocytes, along with atrial and ventricular non-myocytes, simultaneously from a single mouse heart. Also provided are the details for optimal cell-specific culturing methods, which enhance cell viability and function. This protocol aims not only to expedite the process of adult murine cardiac cell isolation, but also to increase the yield and viability of cells for investigations of atrial and ventricular cardiac cells.

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Section: Discussionmentioning

confidence: 99%

Simultaneous Isolation and Culture of Atrial Myocytes, Ventricular Myocytes, and Non-Myocytes from an Adult Mouse Heart

Blackwood

Bilal

Azizi

et al. 2020

JoVE

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show abstract

“…The protocol described above for calcium reintroduction into the isolated ventricular myocyte cultures was designed to limit cardiac myocyte death via inappropriate calcium influx via store operated calcium channels. It should be noted that the stepwise calcium reintroduction should not be performed for isolated atrial myocyte cultures, as this will promote cell death during short-and long-term culture 12 . For further precaution, the perfusion and digestion buffers used here include the cardiac muscle contraction inhibitor butanedione monoxime (BDM) to avoid hypercontraction of isolated myocytes, as well as the calcium paradox, both of which impact myocyte viability 26 .…”

Section: Discussionmentioning

confidence: 99%

Simultaneous Isolation and Culture of Atrial Myocytes, Ventricular Myocytes, and Non-Myocytes from an Adult Mouse Heart

Blackwood¹,

Bilal²,

Azizi³

et al. 2020

JoVE

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The isolation and culturing of cardiac myocytes from mice has been essential for furthering the understanding of cardiac physiology and pathophysiology. While isolating myocytes from neonatal mouse hearts is relatively straightforward, myocytes from the adult murine heart are preferred. This is because compared to neonatal cells, adult myocytes more accurately recapitulate cell function as it occurs in the adult heart in vivo. However, it is technically difficult to isolate adult mouse cardiac myocytes in the necessary quantities and viability, which contributes to an experimental impasse. Furthermore, published procedures are specific for the isolation of either atrial or ventricular myocytes at the expense of atrial and ventricular non-myocyte cells. Described here is a detailed method for isolating both atrial and ventricular cardiac myocytes, along with atrial and ventricular non-myocytes, simultaneously from a single mouse heart. Also provided are the details for optimal cell-specific culturing methods, which enhance cell viability and function. This protocol aims not only to expedite the process of adult murine cardiac cell isolation, but also to increase the yield and viability of cells for investigations of atrial and ventricular cardiac cells.

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“…Right and left atrial myocytes were isolated as previously described ( Egom et al, 2015 ; Jansen et al, 2018 ; Jansen and Rose, 2019 ). Briefly, mice were anaesthetized using isoflurane inhalation and then sacrificed by cervical dislocation.…”

Section: Methodsmentioning

confidence: 99%

Regional and temporal progression of atrial remodeling in angiotensin II mediated atrial fibrillation

Jansen

McRae²,

Mackasey³

et al. 2022

Front. Physiol.

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Atrial fibrillation (AF) is associated with electrical and structural remodeling in the atria; however, the regional and temporal progression of atrial remodeling is incompletely understood. The objective of this study was to investigate the regional and temporal progression of atrial remodeling leading to changes in AF susceptibility in angiotensin II (Ang II) mediated hypertension. Mice were infused with Ang II for 3, 10 or 21 days. AF susceptibility and atrial electrophysiology were studied in vivo using intracardiac electrophysiology. Right and left atrial myocyte electrophysiology was studied using patch-clamping. Atrial fibrosis was assessed histologically. P wave duration and atrial effective refractory period increased progressively from 3 to 21 days of Ang II. AF susceptibility tended to be increased at 10 days of Ang II and was elevated at 21 days of Ang II. Left, but not right, atrial AP upstroke velocity and Na+ current were reduced at 10 and 21 days of Ang II. Left atrial action potential (AP) duration increased progressively from 3 to 21 days of Ang II due to reductions in repolarizing K+ current. Right atrial AP prolongation was increased only after 21 days of Ang II. Left and right atrial fibrosis developed progressively from 3 to 21 days, but increases were larger in the left atrium. In conclusion, Ang II mediated atrial electrical and structural remodeling develop earlier and more extensively in the left atrium compared to the right atrium, providing insight into how atrial remodeling leads to enhanced AF susceptibility in Ang II mediated hypertension.

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Isolation of Atrial Myocytes from Adult Mice

Cited by 7 publications

References 21 publications

Simultaneous Isolation and Culture of Atrial Myocytes, Ventricular Myocytes, and Non-Myocytes from an Adult Mouse Heart

Simultaneous Isolation and Culture of Atrial Myocytes, Ventricular Myocytes, and Non-Myocytes from an Adult Mouse Heart

Simultaneous Isolation and Culture of Atrial Myocytes, Ventricular Myocytes, and Non-Myocytes from an Adult Mouse Heart

Regional and temporal progression of atrial remodeling in angiotensin II mediated atrial fibrillation

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