Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Electron-microscopic studies on reaggregate cultures of vascular smooth muscle cells from normotensive and spontaneously hypertensive rats

  • 19 Accesses

  • 4 Citations


Vascular smooth muscle cells were taken from the aortae of the WKY (normotensive) and SHR (spontaneously hypertensive) strains of rat by enzymatic dispersion and put into reaggregate culture. Initially the cells became individual spheroids having average diameters of 10 μm and surfaces that were either rough or smooth. The cells were far more complex than they appeared on their surfaces; after one day in culture, there was considerable internal variation in these cells. All the cells, whether WKY or SHR, lost the bulk of their cytoplasmic contents (including myofilaments, many mitochondria, and vesicular structures) in the early stages of culture and eventually became flattened. After 14 days in culture, these modified cells collected to form reaggregates that were commonly roughly spherical and several hundred μm in diameter. These reaggregates consisted of peripheral regions made up of several layers of flattened cells overlying cores formed by glia-like networks of cells similar in cytological appearance to the cells at the periphery. The meshes formed in this way contained cellular debris derived from dead cells or extrusion of cellular contents. It appears that SHR cells are quicker to form reaggregates than are WKY cells. Yet the SHR cells retained a rounded conformation after five days, whereas the WKY cells were more flattened and formed a more discrete aggregate at this stage of culture. However, by the fourteenth day of culture, differences between the two cell strains were not so pronounced, as far as could be judged by observations made with scanning and transmission electron microscopy. Both WKY and SHR cells at 14 days appeared highly secretory, possessing large Golgi systems as well as numerous ER cisternae and mitochondria. SHR cells produced greater amounts of connective tissue at all stages of culture than did WKY cells, indicating that a similar difference may contribute to the hypertension which develops naturally in situ in SHR animals.

This is a preview of subscription content, log in to check access.


  1. Bierman EL, Stein O, Stein Y (1974) Lipoprotein uptake and metabolism by rat aortic smooth muscle cells in tissue culture. Circ Res 35:136–150

  2. Brown BG, Mahley R, Assmann G (1976) Swine aortic smooth muscle in tissue culture. Some effects of purified swine lipoproteins on cell growth and morphology. Circ Res 39:415–424

  3. Buck RC (1977) Organ cultures of rat aorta: a scanning and transmission electron microscopic study. Exp Molec Pathol 26:260–276

  4. Campbell GR, Chamley JH (1975) Thick filaments in vertebrate smooth muscle. Cell Tissue Res 156:201–216

  5. Campbell GR, Chamley JH (1976) Ultrastructure of differentiating vascular smooth muscle. In: Vascular neuroeffector mechanisms, 2nd Int Symp. Karger, Basel, pp 1–9

  6. Chamley-Campbell J, Campbell GR, Ross R (1979) The smooth muscle cell in culture. Physiol Rev 59:1–61

  7. Clark JM, Glagov S (1976) Evaluation and publication of scanning electron micrographs. Science 192:1360–1361

  8. Coltoff-Schiller B, Goldfischer S, Wolinsky H, Factor SM (1976) Lipid accumulation in human aortic smooth muscle cell lysosomes. Am J Pathol 83:39–44

  9. Devine CE, Somlyo AP (1971) Thick filaments in vascular smooth muscle. J Cell Biol 49:636–649

  10. Devine CE, Somlyo AV, Somlyo AP (1973) Sarcoplasmic reticulum and mitochondria as cation accumulation sites in smooth muscle. Phil Trans Roy Soc Lond B 265:17–23

  11. Forbes MS (1971) Fine structure of the stellate cell in the pars distalis of the lizard, Anolis carolinensis. J Morphol 136:227–246

  12. Garfield RE, Chacko S, Blose S (1975) Phagocytosis by muscle cells. Lab Invest 33:418–427

  13. Gimbrone MA, Cotran RS (1975) Human vascular smooth muscle in culture. Growth and ultrastructure. Lab Invest 33:16–27

  14. Harder DR, Sperelakis N (1979) Action potential generation in reaggregates of rat aortic smooth muscle cells in primary culture. Blood Vessels 16:186–201

  15. Harrison CJ, Allen TD (1979) Cell surface morphology after trypsinization depends on initial cell shape. Differentiation 15:61–66

  16. Hinek A, Thyberg J (1977) Electron microscopic observations on the formation of elastic fibers in primary cultures of aortic smooth muscle cells. J Ultrastruct Res 60:12–20

  17. Ichijima K (1969) Morphological studies on the peripheral small arteries of spontaneously hypertensive rats. Jpn Circ J 33:785–813

  18. Jarmolych JA, Daoud S, Landau J, Fritz E, McElvene E (1968) Aortic media explants. Cell proliferation and production of mucopolysaccharides, collagen and elastic tissue. Exp Mol Pathol 9:171–188

  19. Larsen WJ (1977) Structural diversity of gap junctions. A review. Tissue and Cell 9:373–394

  20. Lee RMKW, Garfield RE, Forrest JB, Daniel EE (1979) The effects of fixation, dehydration and critical point drying on the size of cultured smooth muscle cells. Scanning Electron Microscopy/1979/III, 439–448

  21. Limas C, Westrum B, Limas CJ (1980) The evolution of vascular changes in the spontaneously hypertensive rat. Am J Pathol 98:357–384

  22. Lompré AM, Poggioli J, Vassort G (1979) Maintenance of fast Na-channels during primary culture of embryonic chick heart cells. J Mol Cell Cardiol 11:813–825

  23. Mayne R, Vail MS, Blose S, Chacko S (1976) Aortic smooth muscle cells grown in culture synthesize both type I and type III collagen. J Cell Biol 70:122a

  24. McLean MJ, Sperelakis N (1977) Electrophysiological recordings from spontaneously contracting reaggregates of cultured vascular smooth muscle cells from chick embryos. Exp Cell Res 104:309–318

  25. McLean MJ, Pelleg A, Sperelakis N (1979) Electrophysiological recordings from spontaneously contracting reaggregates of cultured smooth muscle cells from guinea pig vas deferens. J Cell Biol 80:539–552

  26. Moskalewski S, Konwinski M, Hinek A (1976) In vitro elastic fiber formation by aggregated aortic cells of newborn rabbit. Anat Embryol 150:113–122

  27. Narayanan AS, Sandberg LB, Ross R, Layman DL (1976) The smooth muscle cell. III. Elastin synthesis in arterial smooth muscle cell culture. J Cell Biol 68:411–419

  28. Okamoto K, Aoki K (1963) Development of a strain of spontaneously hypertensive rats. Jpn Circ J 27:282–293

  29. Rice RV, McManus GM, Devine CE, Somlyo AP (1971) Regular organization of thick filaments in mammalian smooth muscle. Nature 231:242–243

  30. Ross R (1971) The smooth muscle cell. II. Growth of smooth muscle in culture and formation of elastic fibers. J Cell Biol 50:172–186

  31. Shimada Y, Fischman DA (1976) Cardiac cell aggregation by scanning electron microscopy. In: Lieberman M, Sano T (eds) Developmental and physiological correlates of cardiac muscle. Raven Press, New York, pp 81–102

  32. Sperelakis N (1978) Cultured heart cell reaggregate model for studying cardiac toxicology. Environ Health Perspect 26:243–267

  33. Sperelakis N, McLean MJ (1978) Electrical properties of cultured heart cells. In: Kobayashi T, Ito Y, Rona G (eds) Recent advances in studies on cardiac structure and metabolism, Vol 12, Cardiac adaptation. University Park Press, Baltimore, pp 645–666

  34. Venable JH, Coggeshall R (1965) A simplified lead citrate stain for use in electron microscopy. J Cell Biol 25:407–408

  35. Warshaw DM, Mulvany MJ, Halpern W (1979) Mechanical and morphological properties of arterial resistance vessels in young and old spontaneously hypertensive rats. Circ Res 45:250–259

  36. Wight TN, Cooke PH, Smith SC (1977) An electron microscopic study of pigeon aorta cell cultures. Cytodifferentiation and intracellular lipid accumulation. Exp Molec Pathol 27:1–18

  37. Yoneda S (1978) Fine surface structure of heart cells in culture — in comparison with the changes in heart cells due to changes of the substratum and L-cell. Jpn Circ J 42:1093–1101

  38. Zelcer E, Sperelakis N (1981) Angiotensin induction of active responses in cultured reaggregates of rat aortic smooth muscle cells (Submitted for publication)

Download references

Author information

Correspondence to M. S. Forbes Ph.D..

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Yoneda, S., Forbes, M.S., Zelcer, E. et al. Electron-microscopic studies on reaggregate cultures of vascular smooth muscle cells from normotensive and spontaneously hypertensive rats. Cell Tissue Res. 217, 225–243 (1981).

Download citation

Key words

  • Vascular smooth muscle
  • Spontaneously hypertensive rat
  • Reaggregate cultures
  • Ultrastructure
  • Collagen synthesis