Nonspecific proteases: a new approach to the isolation of adult cardiocytes.
The potent collagenolytic activity of nonspecific proteinases suggested their use as enzymatic agents for the dissociation of single adult heart cells. This was assessed in guinea pig hearts perfused for 1 min with solutions containing hyaluronidase (100-10 000 U/mL), trypsin (100-10 000 U/mL), crude collagenase (100-500 U/mL), or nonspecific protease (0.1-100 U/mL). No rod-shaped cells were observed among the cells isolated with these concentrations of hyaluronidase, trypsin, or crude collagenase. By contrast, 45-80% of the cells released with nonspecific protease (5-10 U/mL) were rod shaped and Ca2+ tolerant. Resting and action potentials recorded from cells dispersed with nonspecific protease were similar to those recorded from cells isolated after prolonged collagenase exposure.
D000200:Action Potentials; D000818:Animals; D002469:Cell Separation / Q000379:methods*; D006168:Guinea Pigs; D006321:Heart / Q000502:physiology; D003012:Microbial Collagenase / Q000494:pharmacology; D009206:Myocardium / Q000166:cytology*; D010447:Peptide Hydrolases / Q000494:pharmacology*
D010447:Peptide Hydrolases; D003012:Microbial Collagenase