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Cerebral microvascular smooth muscle in tissue culture

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Summary

Cerebral endothelium is being studied rather extensively in tissue culture, but no reports are available describing the tissue culture of cerebral microvascular smooth muscle. The present paper describes for the first time the isolation and culture of non-neoplastic mouse cerebral vascular smooth muscle. Microvessels from a dounce homogenate of mouse brain are plated onto plastic culture dishes in Dulbecco’s modified Eagle media plus 20% fetal bovine serum and treated briefly with collagenase. Cells migrate from vessels and proliferate sufficiently to be transferred out of primary culture in 2 to 3 wk. Light microscopy reveals generally broad, polygonal cells that grow collectively in a “hill and valley” pattern. By transmission electron microscopy the cells possess many characteristics of smooth muscle: basal laminas, clusters of pinocytotic vesicles, and bundles of thin filaments. Several ill-defined cell-to-cell junctions are also present. Isoelectric focusing and sodium dodecyl sulfate-electrophoresis of cellular proteins on polyacrylamide gels after pulse labeling cultures with [S-35]methionine demonstrate that these cells actively synthesize a smooth-muscle-specific isoactin, α-actin. The identity of α-actin is confirmed by analysis of NH2-terminal peptides after actin digestion with trypsin and subsequent peptide cleavage with thermolysin. Both their morphology and active synthesis of α-actin strongly suggest that these cells are of smooth-muscle origin. Future studies of their metabolism and interactions with endothelium and astrocytes should provide a better understanding of the cerebral microcirculation.

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Authors and Affiliations

  1. Department of Pathology, The University of Iowa Hospitals and Clinics, 52242, Iowa City, Iowa

    Steven A. Moore, Elizabeth J. Yoder & Michael N. Hart

  2. Department of Biochemistry, The University of Iowa Hospitals and Clinics, 52242, Iowa City, Iowa

    Arthur R. Strauch & Peter A. Rubenstein

  3. Cardiovascular Research Center, The University of Iowa Hospitals and Clinics, 52242, Iowa City, Iowa

    Arthur R. Strauch

Authors
  1. Steven A. Moore
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  2. Arthur R. Strauch
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  3. Elizabeth J. Yoder
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  4. Peter A. Rubenstein
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  5. Michael N. Hart
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Additional information

This work was supported by Veteran’s Administration Research Funds, National Institutes of Health Grant HL 14230 (Arteriosclerosis Specialized Center of Research, sponsored by the National Heart, Lung, and Blood Institute), and Cardiovascular Research Program Project Grant from the National Institutes of Health to the University of Iowa (HL-14388). A. R. S. is a postdoctoral fellow of the National Institute of General Medical Sciences (GM-09020). P. A. R. is an established investigator of the American Heart Association.

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Moore, S.A., Strauch, A.R., Yoder, E.J. et al. Cerebral microvascular smooth muscle in tissue culture. In Vitro 20, 512–520 (1984). https://doi.org/10.1007/BF02619625

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  • DOI: https://doi.org/10.1007/BF02619625

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