Abstract
Smooth muscle cells were enzymatically dispersed from vasa deferentia of adult male guinea pigs (250-400 g). These cells reassociated in vitro to form monolayers and small spherical reaggregates (0.05-0.3 mm in Diam). Within 48 h of being placed in culture, cells in both types of preparation began to contract spontaneously. The contractions were rhythmic and slow. Cells in the monolayers stopped contracting after approximately 1 wk in vitro, but the reaggregates continued to contract spontaneously for at least 3 wk. Electron microscopy of the reaggregates revealed the presence of thick and thin myofilaments. Overshooting action potentials were recorded in many of the cells penetrated (primarily in reaggregates), and were accompanied by visible contractions of the aggregate or monolayer. Quiescent cells could often be excited by intracellularly applied depolarizing and hyperpolarizing (anodal-break) current pulses. The resting potentials had a mean value of -58 +/- 2 mV. The action potentials were usually preceded by a spontaneous depolarization. The action potentials had slow rates of rise (1--4 V/s) which were unaffected by tetrodotoxin (TTX, 1 microgram/ml), a known blocker of fast Na+ -channels. Verapamil (1 microgram/ml) blocked the action potentials. The mean value of input resistance was 6.9 +/- 0.5 M omega (n = 12). These electrophysiological properties are similar to those of intact adult vas deferens smooth muscle cells. Thus, the cultured adult vas deferens smooth muscle cells retain their functional properties in vitro even after long periods.
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Selected References
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- Anderson D. K. Cell potential and the sodium-potassium pump in vascular smooth muscle. Fed Proc. 1976 May 1;35(6):1294–1297. [PubMed] [Google Scholar]
- Burke J. M., Ross R. Collagen synthesis by monkey arterial smooth muscle cells during proliferation and quiescence in culture. Exp Cell Res. 1977 Jul;107(2):387–395. doi: 10.1016/0014-4827(77)90360-3. [DOI] [PubMed] [Google Scholar]
- Chamley J. H., Campbell G. R., Burnstock G. Dedifferentiation, redifferentiation and bundle formation of smooth muscle cells in tissue culture: the influence of cell number and nerve fibres. J Embryol Exp Morphol. 1974 Oct;32(2):297–323. [PubMed] [Google Scholar]
- Chamley J. H., Campbell G. R., McConnell J. D., Gröschel-Stewart U. Comparison of vascular smooth muscle cells from adult human, monkey and rabbit in primary culture and in subculture. Cell Tissue Res. 1977 Feb 14;177(4):503–522. doi: 10.1007/BF00220611. [DOI] [PubMed] [Google Scholar]
- Chamley J. H., Campbell G. R. Mitosis of contractile smooth muscle cells in tissue culture. Exp Cell Res. 1974 Mar 15;84(1):105–110. doi: 10.1016/0014-4827(74)90385-1. [DOI] [PubMed] [Google Scholar]
- Dehaan R. L., Fozzard H. A. Membrane response to current pulses in spheroidal aggregates of embryonic heart cells. J Gen Physiol. 1975 Feb;65(2):207–222. doi: 10.1085/jgp.65.2.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goto K., Millecchia L. L., Westfall D. P., Fleming W. W. A comparison of the electrical properties and morphological characteristics of the smooth muscle of the rat and guinea-pig vas deferens. Pflugers Arch. 1977 Apr 25;368(3):253–261. doi: 10.1007/BF00585204. [DOI] [PubMed] [Google Scholar]
- Gröschel-Stewart U., Chamley J. H., Campbell G. R., Burnstock G. Changes in myosin distribution in dedifferentiating and redifferentiating smooth muscle cells in tissue culture. Cell Tissue Res. 1975 Dec 29;165(1):13–22. doi: 10.1007/BF00222796. [DOI] [PubMed] [Google Scholar]
- Hiraoka M., Hecht H. H. Recovery from hypothermia in cardiac Purkinje fibers: considerations for an electrogenic mechanism. Pflugers Arch. 1973 Mar 5;339(1):25–36. doi: 10.1007/BF00586979. [DOI] [PubMed] [Google Scholar]
- McLean M. J., Sperelakis N. Electrophysiological recordings from spontaneously contracting reaggregates of cultured vascular smooth muscle cells from chick enbryos. Exp Cell Res. 1977 Feb;104(2):309–318. doi: 10.1016/0014-4827(77)90096-9. [DOI] [PubMed] [Google Scholar]
- Noma A., Irisawa H. Electrogenic sodium pump in rabbit sinoatrial node cell. Pflugers Arch. 1974;351(2):177–182. doi: 10.1007/BF00587436. [DOI] [PubMed] [Google Scholar]
- Ross R., Glomset J. A. Atherosclerosis and the arterial smooth muscle cell: Proliferation of smooth muscle is a key event in the genesis of the lesions of atherosclerosis. Science. 1973 Jun 29;180(4093):1332–1339. doi: 10.1126/science.180.4093.1332. [DOI] [PubMed] [Google Scholar]
- Ross R. The smooth muscle cell. II. Growth of smooth muscle in culture and formation of elastic fibers. J Cell Biol. 1971 Jul;50(1):172–186. doi: 10.1083/jcb.50.1.172. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shigenobu K., Schneider J. A., Sperelakis N. Verapamil blockade of slow Na+ and Ca++ responses in myocardial cells. J Pharmacol Exp Ther. 1974 Aug;190(2):280–288. [PubMed] [Google Scholar]
- Somlyo A. P., Somlyo A. V. Vascular smooth muscle. I. Normal structure, pathology, biochemistry, and biophysics. Pharmacol Rev. 1968 Dec;20(4):197–272. [PubMed] [Google Scholar]
- Sperelakis N., McLean M. J. Electrical properties of cultured heart cells. Recent Adv Stud Cardiac Struct Metab. 1976 May 26;12:645–666. [PubMed] [Google Scholar]
- Taylor G. S., Paton D. M., Daniel E. E. Characteristics of electrogenic sodium pumping in rat myometrium. J Gen Physiol. 1970 Sep;56(3):360–375. doi: 10.1085/jgp.56.3.360. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas R. C. Electrogenic sodium pump in nerve and muscle cells. Physiol Rev. 1972 Jul;52(3):563–594. doi: 10.1152/physrev.1972.52.3.563. [DOI] [PubMed] [Google Scholar]