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

Abstract

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.

Zbtb16 increases susceptibility of atrial fibrillation in type 2 diabetic mice via Txnip-Trx2 signaling

“…Second, we used HL-1 cell line to perform experiments in vitro since there was no well-established protocol to culture adult mouse primary atrial myocytes. Although some studies provided probability for culturing [ 52 , 53 ], atrial myocytes cultured in HG condition have not been proved and estimated.…”

Revisiting Cardiac Biology in the Era of Single Cell and Spatial Omics

“…Postmitotic cardiomyocytes constitute only a third of the cells in the adult mammalian heart, yet they are the predominant cardiac cell type due to their large size, which presents challenges for single-cell isolation. 12 Cardiac cells have been extensively studied using scRNA-seq analysis, [13][14][15][16] however cell isolation techniques impose size constraints, particularly in popular droplet-based sequencing methods that exclude large cell types, such as cardiomyocytes. Some nanowell approaches, such as ICell8, 17 have successfully captured intact cardiomyocytes as an alternative targeted approach with limited throughput.…”

“…Cardiac cells have been extensively studied using scRNA-seq analysis, 13–16 however cell isolation techniques impose size constraints, particularly in popular droplet-based sequencing methods that exclude large cell types, such as cardiomyocytes. Some nanowell approaches, such as ICell8, 17 have successfully captured intact cardiomyocytes as an alternative targeted approach with limited throughput.…”

Optimizing Adeno-Associated Virus Serotype 9 for Studies of Cardiac Chamber–Specific Gene Regulation

“…To determine whether the chamber specificity of AAV9-A and AAV9-V was attributed to myocyte-specific expression, atrial and ventricular myocytes and nonmyocytes were isolated from adult mT/mG mouse hearts, as described previously. 5 After injection with AAV9-A or AAV9-V, GFP was most abundantly expressed in atrial myocytes or ventricular myocytes, respectively, whereas being nearly undetectable in atrial and ventricular nonmyocytes (Figure [G]).…”