Isolation of calcium tolerant myocytes from adult rat hearts: Review of the literature and description of a method

Barbara B. Farmer, Michael Mancina, Eric Williams, August M. Watanabe

Research output: Contribution to journalArticle

101 Citations (Scopus)

Abstract

Myocytes have been isolated from adult rat hearts since 1969. The early preparations exhibited the Ca2+ paradox. Over the ensuing years, numerous groups have reported the isolation of Ca2+ tolerant cardiac myocytes. In the present review, detailed comparisons have been made of the yields, viability, and relative Ca2+ tolerance of these different myocyte preparations. The factors to which these investigators attributed the increased Ca2+ tolerance are considered and the current information regarding the mechanism of the Ca2+ paradox is reviewed. A method is given which incorporates several of the modifications described. By this method 40-60% of the ventricular weight was disaggregated into single myocytes within 45 min after the sacrifice of the rats. Viability without further purification was 82 ± 0.7% (n = 35) and Nai+/Ki+ ratios were normal. Upon incubation with 2 mM Ca2+ for 1 hr at 37°C, viability decreased by 6% and ATP and creatine phosphate remained at physiological levels. The preparation is very stable since upon incubation in culture medium containing fetal bovine serum and 1.25 mM free Ca2+ at 25°C for 20 hr, viability decreased only 13% (rod-shaped and trypan blue criteria). The factors which contribute to the quality and Ca2+ tolerance of this preparation are discussed.

Original languageEnglish
Pages (from-to)1-18
Number of pages18
JournalLife Sciences
Volume33
Issue number1
DOIs
StatePublished - Jul 4 1983

Fingerprint

Muscle Cells
Rats
Calcium
Trypan Blue
Phosphocreatine
Cardiac Myocytes
Purification
Culture Media
Adenosine Triphosphate
Research Personnel
Weights and Measures
Serum

ASJC Scopus subject areas

  • Pharmacology

Cite this

Isolation of calcium tolerant myocytes from adult rat hearts : Review of the literature and description of a method. / Farmer, Barbara B.; Mancina, Michael; Williams, Eric; Watanabe, August M.

In: Life Sciences, Vol. 33, No. 1, 04.07.1983, p. 1-18.

Research output: Contribution to journalArticle

Farmer, Barbara B. ; Mancina, Michael ; Williams, Eric ; Watanabe, August M. / Isolation of calcium tolerant myocytes from adult rat hearts : Review of the literature and description of a method. In: Life Sciences. 1983 ; Vol. 33, No. 1. pp. 1-18.
@article{9f1e4e6afae94209bbdfd5b2c68c6076,
title = "Isolation of calcium tolerant myocytes from adult rat hearts: Review of the literature and description of a method",
abstract = "Myocytes have been isolated from adult rat hearts since 1969. The early preparations exhibited the Ca2+ paradox. Over the ensuing years, numerous groups have reported the isolation of Ca2+ tolerant cardiac myocytes. In the present review, detailed comparisons have been made of the yields, viability, and relative Ca2+ tolerance of these different myocyte preparations. The factors to which these investigators attributed the increased Ca2+ tolerance are considered and the current information regarding the mechanism of the Ca2+ paradox is reviewed. A method is given which incorporates several of the modifications described. By this method 40-60{\%} of the ventricular weight was disaggregated into single myocytes within 45 min after the sacrifice of the rats. Viability without further purification was 82 ± 0.7{\%} (n = 35) and Nai+/Ki+ ratios were normal. Upon incubation with 2 mM Ca2+ for 1 hr at 37°C, viability decreased by 6{\%} and ATP and creatine phosphate remained at physiological levels. The preparation is very stable since upon incubation in culture medium containing fetal bovine serum and 1.25 mM free Ca2+ at 25°C for 20 hr, viability decreased only 13{\%} (rod-shaped and trypan blue criteria). The factors which contribute to the quality and Ca2+ tolerance of this preparation are discussed.",
author = "Farmer, {Barbara B.} and Michael Mancina and Eric Williams and Watanabe, {August M.}",
year = "1983",
month = "7",
day = "4",
doi = "10.1016/0024-3205(83)90706-3",
language = "English",
volume = "33",
pages = "1--18",
journal = "Life Sciences",
issn = "0024-3205",
publisher = "Elsevier Inc.",
number = "1",

}

TY - JOUR

T1 - Isolation of calcium tolerant myocytes from adult rat hearts

T2 - Review of the literature and description of a method

AU - Farmer, Barbara B.

AU - Mancina, Michael

AU - Williams, Eric

AU - Watanabe, August M.

PY - 1983/7/4

Y1 - 1983/7/4

N2 - Myocytes have been isolated from adult rat hearts since 1969. The early preparations exhibited the Ca2+ paradox. Over the ensuing years, numerous groups have reported the isolation of Ca2+ tolerant cardiac myocytes. In the present review, detailed comparisons have been made of the yields, viability, and relative Ca2+ tolerance of these different myocyte preparations. The factors to which these investigators attributed the increased Ca2+ tolerance are considered and the current information regarding the mechanism of the Ca2+ paradox is reviewed. A method is given which incorporates several of the modifications described. By this method 40-60% of the ventricular weight was disaggregated into single myocytes within 45 min after the sacrifice of the rats. Viability without further purification was 82 ± 0.7% (n = 35) and Nai+/Ki+ ratios were normal. Upon incubation with 2 mM Ca2+ for 1 hr at 37°C, viability decreased by 6% and ATP and creatine phosphate remained at physiological levels. The preparation is very stable since upon incubation in culture medium containing fetal bovine serum and 1.25 mM free Ca2+ at 25°C for 20 hr, viability decreased only 13% (rod-shaped and trypan blue criteria). The factors which contribute to the quality and Ca2+ tolerance of this preparation are discussed.

AB - Myocytes have been isolated from adult rat hearts since 1969. The early preparations exhibited the Ca2+ paradox. Over the ensuing years, numerous groups have reported the isolation of Ca2+ tolerant cardiac myocytes. In the present review, detailed comparisons have been made of the yields, viability, and relative Ca2+ tolerance of these different myocyte preparations. The factors to which these investigators attributed the increased Ca2+ tolerance are considered and the current information regarding the mechanism of the Ca2+ paradox is reviewed. A method is given which incorporates several of the modifications described. By this method 40-60% of the ventricular weight was disaggregated into single myocytes within 45 min after the sacrifice of the rats. Viability without further purification was 82 ± 0.7% (n = 35) and Nai+/Ki+ ratios were normal. Upon incubation with 2 mM Ca2+ for 1 hr at 37°C, viability decreased by 6% and ATP and creatine phosphate remained at physiological levels. The preparation is very stable since upon incubation in culture medium containing fetal bovine serum and 1.25 mM free Ca2+ at 25°C for 20 hr, viability decreased only 13% (rod-shaped and trypan blue criteria). The factors which contribute to the quality and Ca2+ tolerance of this preparation are discussed.

UR - http://www.scopus.com/inward/record.url?scp=0020585784&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0020585784&partnerID=8YFLogxK

U2 - 10.1016/0024-3205(83)90706-3

DO - 10.1016/0024-3205(83)90706-3

M3 - Article

C2 - 6306371

AN - SCOPUS:0020585784

VL - 33

SP - 1

EP - 18

JO - Life Sciences

JF - Life Sciences

SN - 0024-3205

IS - 1

ER -