Method for isolation of human ventricular myocytes from single endocardial and epicardial biopsies.
The study of adult human ventricular cells has been limited by tissue availability. In this study we describe techniques for the isolation of Ca(2+)-tolerant adult human ventricular cells from both transvenous endomyocardial and epicardial biopsies. Ca(2+)-tolerant cells were obtained from 80% of the biopsies processed. Although the yield of Ca(2+)-tolerant myocytes from either type of biopsy was low (1-5%), myocytes with normal resting potentials and action potentials can be obtained from single biopsy specimens, providing a source of normal human myocytes for electrophysiological study. Resting potentials (Vrest) were recorded in 41 isolated right ventricular endomyocardial cells at 37 degrees C. Sixteen cells were depolarized (Vrest = -26 +/- 13 mV), and 25 cells had normal resting potentials (Vrest = -84 +/- 6 mV). Action potentials were recorded in 16 cells. At a pacing cycle length of 1 s, 4 cells had prolonged action potential duration at 90% (APD90, 718 +/- 26 ms) and 10 cells had normal APD90 (381 +/- 94 ms) compared with those recorded from intact right ventricular septal trabeculae from explanted hearts. Voltage-clamp studies of isolated human ventricular myocytes obtained from these biopsies document the presence of currents previously reported from cells isolated from explanted hearts.
Peeters
G A
GA
Division of Cardiology Surgery, University of Utah Medical Center, Salt Lake City, USA.
Sanguinetti
M C
MC
Eki
Y
Y
Konarzewska
H
H
Renlund
D G
DG
Karwande
S V
SV
Barry
W H
WH
D000818:Animals; D001706:Biopsy; D002452:Cell Count / Q000187:drug effects; D002469:Cell Separation / Q000379:methods*; D003931:Diacetyl / Q000031:analogs & derivatives / Q000494:pharmacology; D004594:Electrophysiology; D004699:Endocardium / Q000473:pathology*; D006168:Guinea Pigs; D006352:Heart Ventricles; D006801:Humans; D009206:Myocardium / Q000473:pathology*; D010496:Pericardium / Q000473:pathology*; D016276:Ventricular Function
C004717:diacetylmonoxime; D003931:Diacetyl