Enrichment of vital adult cardiac muscle cells by continuous silica sol gradient centrifugation.
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.
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient / Q000379:methods*; D002500:Centrifugation, Isopycnic / Q000379:methods*; D003165:Complement System Proteins; D004077:Digoxin / Q000494:pharmacology; D006321:Heart / Q000187:drug effects; D009206:Myocardium / Q000166:cytology*; D011205:Povidone; D051381:Rats; D012822:Silicon Dioxide
D000900:Anti-Bacterial Agents; C016039:Percoll; D004077:Digoxin; D012822:Silicon Dioxide; D003165:Complement System Proteins; D011205:Povidone; D002118:Calcium