Isolation and characterization of adult rat heart cells

Barbara B. Farmer, Robert Harris, Walter W. Jolly, David R. Hathaway, Allan Katzberg, August M. Watanabe, Alan L. Whitlow, H. R. Besch

Research output: Contribution to journalArticle

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Abstract

Adult rat heart muscle cells were isolated after simultaneous perfusion of multiple (two to eight) hearts with buffered salt solutions containing collagenase and hyaluronidase. Yields (35 to 50% of ventricular weight with approximately 70% viability) are quantitatively suitable for metabolic studies. Viability has been determined by the ability of intact cells to exclude trypan blue and the inability of intact cells to oxidize exogenous succinate. Micrographs show that the fine structure of the isolated cells is well ordered. Cell concentrations of glycogen, glucose 6-phosphate, citrate, and various enzymes were similar to those of intact heart. ATP and creatine phosphate concentrations were lower than in whole hearts. Adenosine 3′,5′-monophosphate concentrations were somewhat elevated. Deoxyribonucleic acid was lower than in whole tissue. The isolated cells retain certain metabolic control mechanisms. The uncoupler of oxidative phosphorylation, 2,4-dinitrophenol, increased oxygen consumption severalfold, whereas exogenous ADP had no effect on respiration. Under anaerobic conditions the rates of glucose utilization and lactate production were faster than in the presence of oxygen, indicating retention of the Pasteur effect. The addition of glucose and insulin caused a decrease in oxygen uptake or the Crabtree effect. Exogenously added pyruvate decreased glycolytic flux and produced a pronounced increase in intracellular citrate and glucose 6-phosphate. Isoproterenol stimulated adenylate cyclase activity of the isolated cells at the same concentrations effective with intact heart preparations. Isoproterenol and glucagon caused the activation of phosphorylase. The cells deteriorated as a function of incubation time, as indicated by a decrease in ATP content and a loss of lactate dehydrogenase into the medium. Cell deterioration was greatly accelerated by Ca2+ at concentrations greater than 10-5 m.

Original languageEnglish
Pages (from-to)545-558
Number of pages14
JournalArchives of Biochemistry and Biophysics
Volume179
Issue number2
DOIs
StatePublished - 1977

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Rats
Glucose-6-Phosphate
Oxygen
Isoproterenol
Citric Acid
Adenosine Triphosphate
2,4-Dinitrophenol
Glucose
Phosphorylases
Hyaluronoglucosaminidase
Trypan Blue
Phosphocreatine
Succinic Acid
Collagenases
Glucagon
Pyruvic Acid
Glycogen
L-Lactate Dehydrogenase
Adenylyl Cyclases
Adenosine

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Farmer, B. B., Harris, R., Jolly, W. W., Hathaway, D. R., Katzberg, A., Watanabe, A. M., ... Besch, H. R. (1977). Isolation and characterization of adult rat heart cells. Archives of Biochemistry and Biophysics, 179(2), 545-558. https://doi.org/10.1016/0003-9861(77)90143-6

Isolation and characterization of adult rat heart cells. / Farmer, Barbara B.; Harris, Robert; Jolly, Walter W.; Hathaway, David R.; Katzberg, Allan; Watanabe, August M.; Whitlow, Alan L.; Besch, H. R.

In: Archives of Biochemistry and Biophysics, Vol. 179, No. 2, 1977, p. 545-558.

Research output: Contribution to journalArticle

Farmer, BB, Harris, R, Jolly, WW, Hathaway, DR, Katzberg, A, Watanabe, AM, Whitlow, AL & Besch, HR 1977, 'Isolation and characterization of adult rat heart cells', Archives of Biochemistry and Biophysics, vol. 179, no. 2, pp. 545-558. https://doi.org/10.1016/0003-9861(77)90143-6
Farmer, Barbara B. ; Harris, Robert ; Jolly, Walter W. ; Hathaway, David R. ; Katzberg, Allan ; Watanabe, August M. ; Whitlow, Alan L. ; Besch, H. R. / Isolation and characterization of adult rat heart cells. In: Archives of Biochemistry and Biophysics. 1977 ; Vol. 179, No. 2. pp. 545-558.
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