Objective To establish a stable and efficient enzyme separation method to isolation of SD rat coronary artery smooth muscle cells (CASMCs), and to study on the voltage dependent potassium ion channels (KV).
Methods CASMCs were isolated by a “three-step” acute enzyme separation method and identified by immunofluorescence. KV currents were recorded by the patch-clamp technique.
Results (1) An abundance of CAMSCs with good reactivity were obtained by this method. (2) At the test potential of +60 mV, the KV current average current densities were 34.35 ± 2.53 pA/pF (n = 5). After added 4-AP, the KV current average current densities were down to 3.55 ± 0.47 pA/pF at the same test potential (P < 0.05). According to Boltzmann equation, the half maximal activation voltage was calculated as 23.66 ± 1.03 mV, and the slope value (k) was 14.44 ± 0.94 mV.
Conclusions This method can acquire a large number of CASMCs which can be used on the study of ion channels.