Abstract
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 (HUVEC), useful primary cells for vascular research. Finally, the maintenance, preservation, and characterization of the isolated vascular cells are described.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Vinters HV (1987) Cerebral amyloid angiopathy: a critical review. Stroke 18(2):311–324
Revesz T, Holton JL, Lashley T, Plant G, Rostagno A, Ghiso J, Frangione B (2002) Sporadic and familial cerebral amyloid angiopathies. Brain Pathol 12(3):343–357
Biffi A, Greenberg SM (2011) Cerebral amyloid angiopathy: a systematic review. J Clin Neurol 7(1):1–9. https://doi.org/10.3988/jcn.2011.7.1.1
Levy E, Carman MD, Fernandez-Madrid IJ, Power MD, Lieberburg I, van Duinen SG, Bots GT, Luyendijk W, Frangione B (1990) Mutation of the Alzheimer’s disease amyloid gene in hereditary cerebral hemorrhage, Dutch type. Sci 248(4959):1124–1126
Gudmundsson G, Hallgrimsson J, Jonasson TA, Bjarnason O (1972) Hereditary cerebral hemorrhage with amyloidosis. Brain 95:387):387–387):404
Bjarnadottir M, Nilsson C, Lindstrom V, Westman A, Davidsson P, Thormodsson F, Blondal H, Gudmundsson G, Grubb A (2001) The cerebral hemorrhage-producing cystatin C variant (L68Q) in extracellular fluids. Amyloid 8(1):1–10
Wisniewski HM, Fraçkowiak J, Zòltowska A, Kim KS (1994) Vascular ß-amyloid in Alzheimer’s disease angiopathy is produced by proliferating and deghenerating smooth muscle cells. Amyloid 1:8–16
Wang ZZ, Jensson O, Thorsteinsson L, Vinters HV (1997) Microvascular degeneration in hereditary cystatin C amyloid angiopathy of the brain. APMIS 105(1):41–47
Wisniewski HM, Frackowiak J, Mazur-Kolecka B (1995) In vitro production of ß-amyloid in smooth muscle cells isolated from amyloid angiopathy-affected vessels. Neurosci Lett 183:120–123
Van Nostrand WE, Davis-Salinas J, Saporito-Irwin SM (1996) Amyloid beta-protein induces the cerebrovascular cellular pathology of Alzheimer’s disease and related disorders. Ann N Y Acad Sci 777:297–302
Wilhelmus MM, Otte-Holler I, van Triel JJ, Veerhuis R, Maat-Schieman ML, Bu G, de Waal RM, Verbeek MM (2007) Lipoprotein receptor-related protein-1 mediates amyloid-beta-mediated cell death of cerebrovascular cells. Am J Pathol 171(6):1989–1999. https://doi.org/10.2353/ajpath.2007.070050
Vilhjalmsson DT, Blondal H, Thormodsson FR (2007) Solubilized cystatin C amyloid is cytotoxic to cultured human cerebrovascular smooth muscle cells. Exp Mol Pathol 83(3):357–360. https://doi.org/10.1016/j.yexmp.2007.09.002
Reynolds MR, Singh I, Azad TD, Holmes BB, Verghese PB, Dietrich HH, Diamond M, Bu G, Han BH, Zipfel GJ (2016) Heparan sulfate proteoglycans mediate Abeta-induced oxidative stress and hypercontractility in cultured vascular smooth muscle cells. Mol Neurodegener 11:9. https://doi.org/10.1186/s13024-016-0073-8
Nicolakakis N, Hamel E (2011) Neurovascular function in Alzheimer’s disease patients and experimental models. J Cereb Blood Flow Metab 31(6):1354–1370. https://doi.org/10.1038/jcbfm.2011.43
Zipfel GJ, Han H, Ford AL, Lee JM (2009) Cerebral amyloid angiopathy: progressive disruption of the neurovascular unit. Stroke 40(3 Suppl):S16–S19. https://doi.org/10.1161/STROKEAHA.108.533174
Mulder AB, Blom NR, Smit JW, Ruiters MH, van der Meer J, Halie MR, Bom VJ (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(5):399–411
Van Nostrand WE, Rozemuller AJM, Chung R, Cotman CW, Saporito-Irwin SM (1994) Amyloid ß-protein precursor in cultured leptomeningeal smooth muscle cells. Amyloid 1:1–7
Acknowledgments
We would like to thank The Icelandic Research Council and Heilavernd (The Icelandic HCHWA-I Foundation) for their support.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Thormodsson, F.R., Olafsson, I.H., Vilhjalmsson, D.T. (2018). Preparation and Culturing of Human Primary Vascular Cells. In: Sigurdsson, E., Calero, M., Gasset, M. (eds) Amyloid Proteins. Methods in Molecular Biology, vol 1779. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7816-8_21
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7816-8_21
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7815-1
Online ISBN: 978-1-4939-7816-8
eBook Packages: Springer Protocols