Abstract
In vitro models utilizing human mesenchymal cells isolated from normal and pathological tissue have proven very useful for studies of wound repair and fibrosis. We have been studying the pathogenesis of intestinal fibrosis and, over the course of 20 yr, have refined techniques for the isolation, culture, and characterization of smooth muscle cells of the human intestinal muscularis propria (1). These methodologies have been adapted for isolation of similar cell types from liver, gallbladder, and blood vessels (2). In this chapter, we describe techniques for the isolation and culture of these cells (procedure A), for the quantitation of procollagen secretion (procedure B), and for the screening of isolates for the expression of smooth muscle-specific cytoskeletal proteins (procedure C).
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Graham, M. F., Diegelmann, R. F., Elson, C. O., Bitar, K. N., and Ehrlich, H. P. (1984) Isolation and culture of human intestinal smooth muscle cells. Proc. Soc. Exp. Biol. Med. 176, 503–507.
Sanyal, A. J., Contos, M. J., Yager, D., Zhu, Y., Willey, A., and Graham, M. F. (1998) Development of pseudointima and stenosis after transjugular intrahepatic portosystemic shunts: characterization of cell phenotype and function. Hepatology 28, 22–31.
Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.
Graham, M. F., Willey, A., Adams, J., Yager, D., and Diegelmann, R. F. (1996) Interleukin 1 beta down-regulates collagen and augments collagenase expression in human intestinal smooth muscle cells. Gastroenterology 110, 344–350.
Graham, M. F., Willey, A., Adams, J., and Diegelmann, R. F. (1995) Corticosteroids increase procollagen gene expression, synthesis, and secretion by human intestinal smooth muscle cells. Gastroenterology 1091, 454–461.
Graham, M. F., Willey, A., Adams, J., Yager, D., and Diegelmann R. F. (1995) Role of ascorbic acid in procollagen expression and secretion by human intestinal smooth muscle cells. J. Cell Physiol. 162, 225–233.
Rosenblat, G., Willey, A., Zhu, Y. N., Jonas, A., Diegelmann, R. F., Neeman, I., and Graham, M. F. (1999) Palmitoyl ascorbate: selective augmentation of procollagen mRNA expression compared with L-ascorbate in human intestinal smooth muscle cells. J. Cell Biochem. 73, 312–320.
Graham, M. F., Gluck, U., Shah, M., and Ben-Ze’ev, A. (1994) β-tropomyosin and α-actin are phenotypic markers for human intestinal smooth muscle cells in vitro. Mol. Cell. Diff. 2(1), 45–60.
Graham, M. F., Bryson, G. R., and Diegelmann, R. F. (1990) Transforming growth factor-β1 selectively augments collagen synthesis by human intestinal smooth muscle cells. Gastroenterology 99, 447–453.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Graham, M., Willey, A. (2003). Isolation, Culture, and Characterization of Human Intestinal Smooth Muscle Cells. In: DiPietro, L.A., Burns, A.L. (eds) Wound Healing. Methods in Molecular Medicine™, vol 78. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-332-1:417
Download citation
DOI: https://doi.org/10.1385/1-59259-332-1:417
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-0-89603-999-5
Online ISBN: 978-1-59259-332-3
eBook Packages: Springer Protocols