Preparation of Cultured Human Vascular Cells

  • Ingvar H. Olafsson
  • Dadi Th. Vilhjalmsson
  • Finnbogi R. ThormodssonEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 849)


Cerebral amyloid angiopathy (CAA) results from amyloid accumulation within arteries of the cerebral cortex and leptomeninges. This condition is age-related, especially prevalent in Alzheimer’s disease (AD), and the main feature of certain hereditary disorders (i.e., HCHWA-I). The vascular smooth muscle cells (VSMCs) appear to play a vital role in the development of CAA, which makes them well suited as an experimental model to study the disease and screen for possible remedies. We describe two different methods for isolating and culturing human VSMCs. First, using the human umbilical cord as an easy source of robust cells, and secondly, using brain tissue that provides the proper cerebral VSMCs, but is more problematic to work with. The umbilical cord also provides human umbilical vascular endothelial cells (HUVECs), useful primary cells for vascular research. Finally, the maintenance, preservation, and characterization of the isolated vascular cells are described.

Key words

Amyloid Cell culture Human Cystatin C Human umbilical vascular endothelial cells Vascular smooth muscle cells Umbilical cord Cerebral amyloid angiopathy HCHWA-I Immunostaining Leptomeninges Cerebral blood vessels 



We would like to thank The Icelandic Research Council and Heilavernd (The Icelandic HCHWA-I Foundation) for their support.


  1. 1.
    Vinters, H. V. (1987) Cerebral amyloid angiopathy: A critical review. Stroke 18, 311–324.PubMedCrossRefGoogle Scholar
  2. 2.
    Revesz, T., Holton, J. L., Lashley, T., Plant, G., Rostagno, A., Ghiso, J., and Frangione, B. (2002) Sporadic and familial cerebral amyloid angiopathies. Brain Pathol 12, 343–357.PubMedCrossRefGoogle Scholar
  3. 3.
    Levy, E., Carman, M. D., Fernandez-Madrid, I. J., Power, M. D., Lieberburg, I., van Duinen, S. G., Bots, G. T., Luyendijk, W., and Frangione, B. (1990) Mutation of the Alzheimer’s disease amyloid gene in hereditary cerebral hemorrhage, Dutch type. Science 248, 1124–1126.PubMedCrossRefGoogle Scholar
  4. 4.
    Gudmundsson, G., Hallgrimsson, J., Jonasson, T. A., and Bjarnason, O. (1972) Hereditary cerebral hemorrhage with amyloidosis. Brain 95, 387–404.PubMedCrossRefGoogle Scholar
  5. 5.
    Bjarnadottir, M., Nilsson, C., Lindstrom, V., Westman, A., Davidsson, P., Thormodsson, F., Blondal, H., Gudmundsson, G., and Grubb, A. (2001) The cerebral hemorrhage-producing cystatin C variant (L68Q) in extracellular fluids. Amyloid: Int. J. Exp. Clin. Invest. 8, 1–10.Google Scholar
  6. 6.
    Wisniewski, H. M., Fraçkowiak, J., Zòltowska, A., and Kim, K. S. (1994) Vascular ß-amyloid in Alzheimer’s disease angiopathy is produced by proliferating and degenerating smooth muscle cells. Amyloid: Int. J. Exp. Clin. Invest. 1, 8–16.Google Scholar
  7. 7.
    Wang, Z. Z., Jensson, O., Thorsteinsson, L., and Vinters, H. V. (1997) Microvascular degeneration in hereditary cystatin C amyloid angiopathy of the brain. APMIS 105, 41–47.PubMedCrossRefGoogle Scholar
  8. 8.
    Wisniewski, H. M., Frackowiak, J., and Mazur-Kolecka, B. (1995) In vitro production of β-amyloid in smooth muscle cells isolated from amyloid angiopathy-affected vessels. Neuroscience Letters 183, 120–123.PubMedCrossRefGoogle Scholar
  9. 9.
    Van Nostrand, W. E., Davis-Salinas, J., and Saporito-Irwin, S. M. (1996) Amyloid beta-protein induces the cerebrovascular cellular pathology of Alzheimer’s disease and related disorders. Ann N Y Acad Sci 777, 297–302.PubMedCrossRefGoogle Scholar
  10. 10.
    Vilhjalmsson, D. T., Blondal, H., and Thormodsson, F. R. (2007) Solubilized cystatin C amyloid is cytotoxic to cultured human cerebrovascular smooth muscle cells. Experimental and Molecular Pathology 83, 357–360.PubMedCrossRefGoogle Scholar
  11. 11.
    Mulder, A. B., Blom, N. R., Smit, J. W., Ruiters, M. H., van der Meer, J., Halie, M. R., and Bom, V. J. (1995) Basal tissue factor expression in endothelial cell cultures is caused by contaminating smooth muscle cells. Reduction by using chymotrypsin instead of collagenase, Thromb. Res. 80, 399–411.PubMedCrossRefGoogle Scholar
  12. 12.
    Van Nostrand, W. E., Rozemuller, A. J. M., Chung, R., Cotman, C. W., and Saporito-Irwin, S. M. (1994) Amyloid ß-protein precursor in cultured leptomeningeal smooth muscle cells, Amyloid: Int. J. Exp. Clin. Invest. 1, 1–7.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Ingvar H. Olafsson
    • 1
  • Dadi Th. Vilhjalmsson
    • 2
  • Finnbogi R. Thormodsson
    • 3
    Email author
  1. 1.Faculty of Medicine/Department of NeurosurgeryUniversity of Iceland/Landspitali University HospitalReykjavikIceland
  2. 2.Department of Surgery (Colorectal section)Malmö University Hospital (SUS Malmö)MalmöSweden
  3. 3.ValaMed ehf, KIM Medical ParkReykjavikIceland

Personalised recommendations