4.5 Review

Methods for isolating atrial cells from large mammals and humans

Journal

JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY
Volume 86, Issue -, Pages 187-198

Publisher

ELSEVIER SCI LTD
DOI: 10.1016/j.yjmcc.2015.07.006

Keywords

Atrial myocytes; Cell isolation; Human; Large mammal; Dog; Sheep; Pig; Collagenase; Protease; Enzyme

Funding

  1. European-North American Atrial Fibrillation Research Alliance grant of Fondation Leducq (ENAFRA) [07CVD03]
  2. European Network for Translational Research in Atrial Fibrillation (EUTRAF) [261057]
  3. German Federal Ministry of Education and Research through the Atrial Fibrillation Competence Network [01Gi0204]
  4. Deutsche Forschungsgemeinschaft [Do 769/1-1-3]
  5. DZHK (German Centre for Cardiovascular Research)
  6. British Heart Foundation [FS0900226487, FS123429565, PG128929970]
  7. British Heart Foundation [PG/12/89/29970, FS/14/4/30532, FS/12/34/29565, FS/09/002/26487] Funding Source: researchfish

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The identification of disturbances in the cellular structure, electrophysiglogy and calcium handling of atrial cardiomyocytes is crucial to the understanding of common pathologies such as atrial fibrillation. Human right atrial specimens can be obtained during routine cardiac surgery and may be used for isolation of atrial myocytes. These samples provide the unique opportunity to directly investigate the effects of human disease on atrial myocytes. However, atrial myocytes vary greatly between patients, there is little if any access to truly healthy controls and the challenges associated with assessing the in vivo effects of drugs or devices in man are considerable. These issues highlight the need for animal models. Large mammalian models are particularly suitable for this purpose as their cardiac structure and electrophysiology are comparable with humans. Here, we review techniques for obtaining atrial cardiomyocytes. We start with background information on solution composition. Agents shown to increase viable cell yield will then be explored followed by a discussion of the use of tissue-dissociating enzymes. Protocols are detailed for the perfusion method of cell isolation in large mammals and the chunk digest methods of cell isolation in humans. (C) 2015 Elsevier Ltd. All rights reserved.

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