Evidence map›Paper›PMID 40610851›Full record

ArticleBMC cardiovascular disorders2025

An optimized Langendorff-free isolating method and electrophysiology studies for adult mouse atrioventricular node cells.

Yuan Chen, Jiaming Zhang, Jiu Pu, Ting Li, Jinfang Yu, Bifang Mai, Shuanglun Xie, Jingdong Li

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Article in BMC cardiovascular disorders, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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5 · Who and what money

Authors and funding

8 authors.

Yuan ChenDepartment of Cardiology, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, China.
Jiaming ZhangDepartment of Cardiology, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, China.
Jiu PuDepartment of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Ting LiDepartment of Cardiology, Wuhan Central Hospital, Wuhan, China.
Jinfang YuDepartment of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Bifang MaiDepartment of Cardiology, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, China.
Shuanglun XieDepartment of Cardiology, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, China. xieshuanglun@mail2.sysu.edu.cn.
Jingdong LiDepartment of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. jingdong-li@mail.hust.edu.cn.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

backgroundThe atrioventricular node (AVN) plays a critical role in coordinating the sequential activation and contraction of the heart's chambers by transmitting electrical impulses from the sinoatrial node (SAN) to the ventricles via the His-Purkinje system. Besides its primary function, the AVN can generate intrinsic pacemaker activity when the SAN fails, and it serves as an important pharmacological target for controlling ventricular rate in the case of atrial arrhythmias. Despite its clinical significance, detailed electrophysiological studies of the AVN have been challenging due to difficulties in isolating viable AVN cells. The dense cellular network and complex structure of the AVN hinder enzymatic digestion, often leading to low yield and poor cell viability. Traditional isolation methods-such as adapting SAN cell protocols or employing the Langendorff perfusion technique-are limited by inadequate enzymatic penetration and preferential perfusion of the ventricular region, which significantly hampers the yield and viability of AVN cells for electrophysiological studies. Therefore, it is necessary to develop an optimized method to isolate high-quality AVN cells.

methodsA refined method that does not rely on the Langendorff technique was used to isolate AVN cells from adult mice. Immunofluorescent imaging was used to confirm the presence of HCN4-positive cells. Patch clamp techniques were employed to record action potentials and ionic currents in AVN cells. Intracellular Ca

resultsWe have developed an improved non-Langendorff perfusion technique that combines targeted enzymatic digestion with enhanced perfusion of the AVN region. By cannulating and ligating the aorta and packing the perfusion cannula tip with gauze, we achieved uniform enzyme distribution throughout the AVN area. This innovation results in a high yield of viable AVN cells that maintain their electrophysiological properties, making them suitable for advanced analyses such as patch-clamp recordings and calcium transient measurements.

conclusionsOur method provides a strong platform for investigating the physiological and pathological roles of the AVN. This approach has the potential to aid in the development of novel therapeutic strategies for atrioventricular conduction disorders.

Indexed as

Action PotentialsAtrioventricular NodeCell SeparationElectrophysiologic Techniques, CardiacMyocytes, CardiacAnimalsCalcium SignalingMaleMiceMice, Inbred C57BLPatch-Clamp TechniquesAtrioventricular conduction ElectrophysiologyCalcium transientMouse atrioventricular node cell isolationPatch clamp

Identifiers

PMID40610851
PMCPMC12226838

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.