Myocytes from atria of adult guinea-pigs were isolated by means of a previously described enzyme perfusion with some modifications.
The problem of Ca-intolerance of the dispersed cells was circumvented by (i) avoiding cooling of the cells below 25 °C and (ii) increasing the Ca concentration slowly already during the enzyme perfusion.
Isolated atrial myocytes were taken in long-term cell culture. Under this condition they become spherical within about 2 days. The rounded stage (cardioballs), which is found in the cultures for a period of ca. 10 days is highly suited for electrophysiological studies using the different recording configurations of the patch clamp technique, including ‘tight-seal whole-cell recording’ with simultaneous cell dialysis.
Key wordssingle heart cell cardioball cell culture whole-cell recording
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