A combined Langendorff-injection technique for simultaneous isolation of single cardiomyocytes from atria and ventricles of the rat heart

Autor: Tatiana Myachina, Xenia Butova, Anastasia Khokhlova
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Single cell mechanics
Contraction (grammar)
CELL ISOLATION
Clinical Biochemistry
Calcium imaging
010501 environmental sciences
01 natural sciences
Sarcomere
ANIMAL EXPERIMENT
ISOLATED HEART
Extracellular matrix
VENTRICULAR FREE WALL
HEART RIGHT VENTRICLE
Interventricular septum
HEART ATRIUM
PRIORITY JOURNAL
INJECTION TECHNIQUE
HEART VENTRICLE WALL
0303 health sciences
Chemistry
Medical Laboratory Technology
medicine.anatomical_structure
Cardiomyocyte isolation
Protocol Article
CELL SUSPENSION
SINGLE CELL MECHANICS
CALCIUM IMAGING
Perfusion
ATRIUM
Science
INTERVENTRICULAR SEPTUM
Connective tissue
LANGENDORFF-INJECTION
CALCIUM
HEART VENTRICLE
Injection technique
03 medical and health sciences
ADULT
CARDIAC MUSCLE CELL
ANIMAL TISSUE
medicine
CARDIOMYOCYTE ISOLATION
NONHUMAN
ARTICLE
030304 developmental biology
0105 earth and related environmental sciences
Langendorff-based perfusion
Atrium (architecture)
INTRACEREBROVENTRICULAR DRUG ADMINISTRATION
Ventricular free wall
LANGENDORFF-BASED PERFUSION
HEART LEFT VENTRICLE
COLLAGEN
Atrium
HEART PERFUSION
SARCOMERE
ANIMAL CELL
Biophysics
RAT
Langendorff-injection
INTRAATRIAL DRUG ADMINISTRATION
Zdroj: MethodsX
MethodsX, Vol 8, Iss, Pp 101189-(2021)
Popis: Single cardiomyocytes are widely used for investigations of the cellular and molecular mechanisms of regulation and modulation of cardiac performance. Intact cardiomyocytes allow one to study in detail cell function avoiding the effects of extracellular matrix and neighboring cells. The most established protocols of cardiomyocyte isolation are based on the isolated heart perfusion using a Langendorff-apparatus or on intraventricular perfusion using a syringe. However, the yield of single cardiomyocytes obtained by these methods may be low due to the cell injury following non-uniform enzyme digestion of connective tissue in different heart chambers. Moreover, isolation of atrial cardiomyocytes is challenging because of their small size and complex geometric shape. Here we present a new protocol for simultaneous isolation of high quality cardiomyocytes from the atria, ventricular free walls and interventricular septum. The protocol is based on the combination of the Langendorff perfusion method with the intraventricular and intra-atrial injection technique taking into account the collagen content variation between the different heart chambers. Obtained cells demonstrate rod-shaped morphology, a clear and regular sarcomere striation pattern and rat-specific frequency-dependence of contraction and calcium transient parameters. Our protocol provides gentle cell isolation that increases the yield of single cardiomyocytes suitable for biophysical researches .
Graphical abstract Image, graphical abstract
Databáze: OpenAIRE
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