Isolation and Contractile Properties of Single Smooth Muscle Cells from Guinea Pig Taenia Caeci

Abstract

A method for the isolation of single smooth muscle cells from guinea pig taenia caeci is described. The single cells were prepared by digestion with 0.3% collagenase and 0.6% trypsin inhibitor in Ca²⁺-free solution. This procedure produced a high yield of intact cells. Most cells obtained by this procedure were relaxed and showed large contractile responses to excitatory stimulus. Maximal responses of the single cells to acetylcholine (ACh), histamine, and high K⁺ were attained within 20sec, and the per cent decrease in cell length was 30-40%. Times for maximal responses of the single cells were shorter than those of the muscle strips. Single cells exhibited a dose-dependent graded response to calcium under depolarized conditions, ACh, histamine, and high K⁺, and a voltage- and duration-dependent response to electrical stimulation. The ED₅₀s of ACh in the single cells and in the muscle strips were about 2×10^-8 and 1.5×10^-7M, respectively. The muscle strips had a lower sensitivity than the single cells to ACh. The generally smooth surface of the relaxed cell contrasted with the numerous evaginations present on the fully contracted cell. I believe that single smooth muscle cells isolated from guinea pig using my technique are, at present, better for physiological and pharmacological studies than are cells isolated using other techniques.