Summary
Maximizing the yield of viable cardiomyocytes suitable for primary cultures is important in the processing of small human myocardial biopsies. The purpose of this study was to establish the conditions of collagenase and trypsin digestion of ventricular myocardium to obtain optimal yields of viable cardiomyocytes. Our results showed that calcium in the digestion solution was toxic to the cells. EDTA chelated calcium in the digestion solution and inhibited collagenase activity. Trypsin increased collagenase activity, especially in the presence of EDTA. The combination of trypsin and collagenase with or without EDTA was more effective than either enzyme alone in increasing cardiomyocyte yield and viability and in minimizing cardiomyocyte damage. By repeating the digestion of a 5 to 20-mg myocardial biopsy, sufficient numbers of viable cardiomyocytes can be obtained for primary culture.
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Li, RK., Tumiati, L.C., Weisel, R.D. et al. Isolation of cardiomyocytes from human myocardium for primary cell culturing. Journal of Tissue Culture Methods 15, 147–154 (1993). https://doi.org/10.1007/BF02388268
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DOI: https://doi.org/10.1007/BF02388268