Dispersion and isolation of beating cells from adult rat heart

Glick, Melvin R.; Burns, Alvin C.; Reddy, William J.

doi:10.1016/0003-2697(74)90329-7

Cited by 93 publications

(23 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Single isolated myocytes were prepared from the ventricles of adult rats, guinea pigs, and bovine hearts by enzymatic dissociation (Glick, Burns & Reddy, 1974;Dow, Hardig & Powell, 1981). The ventricular myocytes of the rat and guinea pig hearts were enzymatically isolated by retrograde perfusion of the aorta, whereas the myocytes of bovine hearts were isolated by incubat- Each magnitude and phase function consists of two superimposed plots, the lower of which is the uncompensated response of the electrode.…”

Section: Methodsmentioning

confidence: 99%

Linear electrical properties of isolated cardiac cells

1984

View full text Add to dashboard Cite

A frequency domain equivalent circuit analysis of isolated ventricular cells indicated the presence of an internal membrane structure which has a total capacitance four- to sixfold larger than the surface membrane. The internal membrane was mainly attributed to the sarcoplasmic reticulum since other morphological studies have shown that its area is many-fold larger than that of the surface membrane. Corresponding estimates from the transverse tubular system indicate an area less than that of the surface; thus this structure is not a likely candidate for the observed internal capacitance. Measurements in hypertonic solutions showed that the access resistance to the internal membrane reversibly increased as the tonicity was elevated. Freeze-fractured electron microscopic studies confirmed that hypertonic solutions increased the volume of transverse tubular system, which thus appears to have little relation to the access resistance. The most probable source of the access resistance is the diadic junction to the sarcoplasmic reticulum, which therefore would electrically couple it to the surface membrane.

show abstract

Section: Methodsmentioning

confidence: 99%

Linear electrical properties of isolated cardiac cells

1984

View full text Add to dashboard Cite

show abstract

“…Single isolated myocytes were prepared from the ventricles of adult guinea pigs and bovine hearts by enzymatic Downloaded from http://ahajournals.org by on March 31, 2019 288 dissociation (Glick et al, 1974;Dow et al, 1981). The ventricular myocytes of the guinea pig hearts were enzymatically isolated by retrograde perfusion of the aorta ("perfusion method"), whereas, the myocytes of bovine hearts were isolated by incubating ventricular tissue chunks in enzyme medium ("chunk method") (Isenberg and Klockner, 1982a).…”

Section: Cell Preparationmentioning

confidence: 99%

Actions of adenosine and isoproterenol on isolated mammalian ventricular myocytes.

Belardinelli¹,

Isenberg²

1983

Circulation Research

181

View full text Add to dashboard Cite

SUMMARY. We investigated the effects of adenosine and isoproterenol on enzymatically dispersed ventricular myocytes from bovine and guinea pig hearts. Inrracellular stimulation of relaxed myocytes with regular srriation patterns and normal resting potential resulted in action potentials with full plateaus accompanied by contractions. Adenosine in concentrations up to 0.2 HIM had no significant effect on any of the action potential parameters or on the basal contractility. In contrast, in the same cells, adenosine effectively antagonized the stimulatory effect of isoproterenol. Isoproterenol (1-10 nM) prolonged the action potentials by 34-41%, displaced the plateau to more positive potentials, and caused a 3-fold increase in the extent of myocyte sarcomere shortening. In the presence of adenosine (5-50 HM), isoproterenol increased the action potential duration by only 8-9%, the shift of the plateau was nearly abolished, and the increase in the extent of myocyte sarcomere shortening was less than 10%. In some of the myocytes, isoproterenol (1-10 nM) induced depolarizing afterpotentials accompanied by aftercontractions. The afterdepolarizations occasionally reached threshold resulting in triggered sustained rhythmic activity. Adenosine (20-50 HM) not only reduced the amplitude of the afterdepolarizarions and aftercontractions, but also abolished the sustained rhythmic activity. We conclude, first, that isolated ventricular myocytes respond to isoproterenol and adenosine; second, that adenosine has no direct effect, but effectively antagonizes the stimulatory actions of isoproterenol; third, that findings are consistent with the ones reported for multicellular ventricular preparations; fourth, that adenosine concentrations required to attenuate the actions of isoproterenol are in the range of adenosine concentrations released by cardiac cells when oxygen availability is limited and/or demand is increased; and fifth, that endogenously released adenosine may modulate the electrophysiological and contractile effects of catecholamines. (Circ Res 53: 287-297, 1983) RECENTLY, adenosine has been recognized to antagonize specifically the inotropic, and some of the electrophysiological and metabolic, effects of catecholamines in ventricular myocardium (Schrader et al., 1977;Dobson, 1978;Baumann et al., 1981, Belardinelli et al., 1982.The observation that adenosine antagonizes the inotropic effect of tyTamine, but not the tyramineinduced release of norepinephrine, has been assumed to indicate that adenosine acts postsynaptically (Schrader et al., 1979). However, adenosine has also been shown to inhibit the stimulationinduced release of norepinephrine from sympathetic nerve terminals in the heart (Hedquist and Fredholm, 1976;Wakade and Wakade, 1978). Consistent with this latter finding, in the dog, adenosine has been shown to reduce the increase in heart rate induced by electric stimulation of the right stellate ganglion (Horn and Lokhandwala, 1981). These observations suggest that the antagonism between adenosine and sympa...

show abstract

“…Percoll leaves the sarcolemmal membrane unaffected. Enriched suspensions of adult cardiocytes are therefore best suited to be used in short-and long-term biochemical and physiological experiments up to 36 hours at 37 ~ In immunological assays determining the cytolytic activity of autoantibodies or cytotoxic T-cells, adult cardiocytes with an intact sarcolemmal membrane are superior (17) to trypsinized cells (9,12,27) that have lost membrane antigens by digestion or to cells that have altered their properties in long-termin culture (25).…”

Section: Discussionmentioning

confidence: 99%

“…Several methods for the isolation of vital adult cardiocytes have been described including treatment with enzymes following mechanical disruption (12,27) or organ perfusion with proteolytic enzymes in calcium free media (1, 3,5,8,13,18,19,23). Even though most investigators used calcium-free media during cell isolation, they usually but not always failed (3,4,6,16) to obtain more than 80 % of the cells morphologically intact.…”

Section: Introductionmentioning

confidence: 99%

Enrichment of vital adult cardiac muscle cells by continuous silica sol gradient centrifugation

Maisch¹

1981

Basic Res Cardiol

View full text Add to dashboard Cite

A major improvement in the isolation of vital adult cardiocytes was achieved by isopycnic preformed continuous silica sol gradient centrifugation after perfusion of the heart with collagenase. Vital rat cardiocytes were enriched to 90-95% vital cells reproducibly and constantly by one- or two-step gradient centrifugations. The isolated cardiocytes were tolerant to calcium concentrations up to 0.03 mmol/l, to diluted human serum, and to human complement. Gentamycin (50 microgram/ml) exerted a cytotoxic effect on myocytes, whereas Penicillium and Streptomycin in concentrations of 50 IU/ml did not induce cytolysis of vital cells. Digoxin 15 ng/ml) decreased the natural decay of myocytes of 20% in 25 hours to 8%. Enriched of vital cardiocytes by silica sol gradient centrifugation following their isolation by perfusion with collagenase may be helpful for investigations depending on a high yield of vital myocardial cells.

show abstract

Dispersion and isolation of beating cells from adult rat heart

Cited by 93 publications

References 11 publications

Linear electrical properties of isolated cardiac cells

Linear electrical properties of isolated cardiac cells

Actions of adenosine and isoproterenol on isolated mammalian ventricular myocytes.

Enrichment of vital adult cardiac muscle cells by continuous silica sol gradient centrifugation

Contact Info

Product

Resources

About