| Abstract: | Single cells were trypsin-dispersed from blood vessels (great vessels near the heart and mesenteric vessels) of 10–20 day chick embryos, and induced to reaggregate into small spheres (0.1–0.5 mm ) either by gyration or by plating on cellophane. Many reaggregates contracted spontaneously or in response to electrical stimulation during culture periods of up to 6 weeks. When the spherical reaggregates were allowed to adhere to a glass substrate, cells emigrated from the spheres to form aprons of monolayered cells which continued to contract. Thick and thin myofilaments (mean diameters of 146 and 65 Å, respectively) were observable in a large fraction of cells studied in electron micrographs. Vascular smooth muscle (VSM) cells were identified in the reaggregates by recording resting potentials of −40 to −60 mV, and by action potential generation. The action potentials were preceded by pacemaker potentials, had slow rates of rise (<20 V/sec), and were insensitive to tetrodotoxin (TTX). Although the action potentials depend on an inward slow current, D-600 did not block the action potentials of the VSM cells. Reaggregates of atrial cells, produced at the same time for comparison, had larger resting potentials (up to −80 mV), less automaticity, fast rates of rise (mean of about 85 V/sec), and complete TTX sensitivity, thus indicating dependence on fast Na+ channels. These findings indicate that identifiable VSM cells can be successfully maintained in primary culture for several weeks, and these cells retain electrical and contractile properties similar to those of smooth muscle cells in intact adult blood vessels. This preparation provides a convenient system for electrophysiological and pharmacological studies of VSM cells. |
