Skip to main content

Cultures of Proliferating Vascular Smooth Muscle Cells from Adult Human Aorta

  • Protocol
Human Cell Culture Protocols

Part of the book series: Methods in Molecular Medicine ((MIMM,volume 2))

Abstract

Abnormal proliferation of human vascular smooth muscle cells (hVSMCs) is a central event in the development of atherosclerosis (13) As a result, there is considerable interest in the establishment of hVSMC cultures as a mode1 of this disease process However, it has been noted in the past (4,5) that hVSMCs, especially when cultured by the enzyme-dispersal technique (hVSMC,ED), grow poorly in culture compared to VSMCs from other species (e.g., rat) This has limited their use for cell-culture studies. We have recently reported that the reduced proliferative capacity of hVSMCED from adult aorta can be attributed to the endogenous production of active TGF-β(6,7). We (68) and others (911) have shown that TGF-β; is a potent inhibtior of smooth muscle cell proliferation. Moreover, recent studies in animal models of atherosclerosis (12) have suggested that TGF-P plays a pivotal role in regulation of vessel wall architecture (13). We have also shown that hVSMCs derived by the alternative method of explanting (hVSMCEX) have a greater proliferative capacity than the hVSMCED (14) In accordance with our hypotheses, the cells grown from explanted tissue did not produce TGF-β; (14).

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 79.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Ross, R. (1986) The pathogenesis of atherosclerosis an update. N Engl J Med 314, 488–500

    Article  PubMed  CAS  Google Scholar 

  2. Ross, R. and Glomset, J. A (1976) The pathogenesis of atherosclerosis I. N. Engl J Med 295, 369–377

    Article  PubMed  CAS  Google Scholar 

  3. Ross, R (1993) The pathogenesis of atherosclerosis. a perspective for the 1990s Nature 362, 80l–809

    Article  Google Scholar 

  4. Dartsch, P, Voisard, R, Baunedel, G, Hofling, B, and Betz, E (1990) Growth characteristics and cytoskeletal organisation of cultured smooth muscle cells from human primary stenosing and restenosing lesions Arteriosclerosis 10, 62–75.

    PubMed  CAS  Google Scholar 

  5. Dartsch, P, Weiss, H.-D, and Betz, E. (1990) Human vascular smooth muscle cells in culture growth characteristics and protein pattern by use of serum-free media supplements. Eur J Cell Biol 51, 285–294.

    PubMed  CAS  Google Scholar 

  6. Kirschenlohr, H. L, Metcalfe, J. C., Weissberg, P L, and Grainger, D. J (1993) Adult human aortlc smooth muscle cells in culture produce active TGF-&#00BD;. Am J Physiol 265, C57l–C576

    Google Scholar 

  7. Grainger, D J, Kirschenlohr, H L, Metcalfe, J C, Weissberg, P L., Wade, D P. and Lawn, R M (1993) Proliferation of human smooth muscle cells promoted by lipoprotein(a) Science 260, 1655–1658

    Article  PubMed  CAS  Google Scholar 

  8. Grainger, D J, Kemp, P. R., Witchell, C M, Weissberg, P L, and Metcalfe, J C (1994) Transforming growth factor &#00DF; decreases the rate of proliferation of rat vascular smooth muscle cells by extending the G, phase of the cell cycle and delays the rise in cychc AMP before entry into M phase Biochem J 299, 227–235

    PubMed  CAS  Google Scholar 

  9. Assoslan, R K and Sporn, M B (1986) Type p transforming growth factor in human platelets release during platelet degranulation and action on vascular smooth muscle cells J Cell Biol 102, 1217–1223

    Article  Google Scholar 

  10. Bjorkerud, S (1991) Effects of transforming growth factor &#00DF;1 on human arterial smooth muscle cells in vitro Arteriosclerosis Thromb 11, 892–902

    CAS  Google Scholar 

  11. Owens, G K, Geisterfer, A A, Yang, Y W, and Kamonya, A (1988) Transforming growth factor &#00DF;-induced growth inhibition and cellular hypertrophy in cultured vascular smooth muscle cells J Cell Biol 107, 77l–780

    Article  Google Scholar 

  12. Grainger, D J, Kemp, P R, Lin, A C, Lawn, R M, and Metcalfe, J C (1994) Activation of transforming growth factor &#00DF; is inhibited in transgenic apolipo protein(a) mice Nature 370, 460–462

    Article  PubMed  CAS  Google Scholar 

  13. Grainger, D J and Metcalfe, J C (1995) A pivotal role for TGF-&#00DF; in athero genesis? Biol Rev Camb Phdos Soc, in press

    Google Scholar 

  14. Kirschenlohr, H L, Metcalfe, J C, Weissberg, P L, and Grainger, D J (1995) Proliferation of human aortlc VSMCs in culture is modulated by active TGF-P Cardiovasc Res 29, 848–855

    PubMed  CAS  Google Scholar 

  15. Majack, R A (1987) Beta-type transforming growth factor specifies organizational behaviour in vascular smooth muscle cell cultures J Cell Biol 105, 465–471

    Article  PubMed  CAS  Google Scholar 

  16. Lemire, J M, Covin, C W, White, S, Giachelli, C M, and Schwartz, S M (1994) Characterization of cloned aortic smooth muscle cells from young rats Am J Pathol 144, 1068–1081.

    PubMed  CAS  Google Scholar 

  17. Jonasson, L, Holm, J, Skalli, O, Gabbiani, G, and Hansson, G K (1985) Expression of class II transplantation antigen on vascular smooth muscle cells in human atherosclerosis J Clan Invest 76, 125–131

    Article  Google Scholar 

  18. Grainger, D J, Witchell, C M., Weissberg, P L., and Metcalfe, J C (1992) Heterogeneity, dedifferentiation and proliferation of primary rat vascular smooth muscle cells and clones J Mol Cell Cardiol 24(suppl. I), 236

    Google Scholar 

  19. Lyons, R. M, Gentry, L E, Purchino, A F, and Moses, H L (1990) Mechanism of activation of latent recombinant transforming growth factor &#00DF;1 by plasmin J Cell Biol 110, 1361–1367

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1996 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Kirschenlohr, H.L., Metcalfe, J.C., Grainger, D.J. (1996). Cultures of Proliferating Vascular Smooth Muscle Cells from Adult Human Aorta. In: Jones, G.E. (eds) Human Cell Culture Protocols. Methods in Molecular Medicine, vol 2. Humana Press. https://doi.org/10.1385/0-89603-335-X:319

Download citation

  • DOI: https://doi.org/10.1385/0-89603-335-X:319

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-335-1

  • Online ISBN: 978-1-59259-586-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics