Abstract
Investigation of cell function is often hampered by the complexity of the tissue context. This problem is circumvented by isolating cells from tissues and analyzing their behavior in culture. Most cell types are cultured as monolayers on planar, rigid Petri dishes, an environment that does not reflect the spatial, three-dimensional cellular environment in vivo. Culture in three-dimensional collagen lattices has been devised to optimize in vitro culture conditions and to provide a more physiologic “in vivo-like” environment. Collagen lattices can easily be manipulated to suit diverse cell types and to provide variable mechanical forces. Cells can be imaged in such surroundings, and gene expression as well as protein production and activity can be monitored.
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
Bell E, Ivarsson B, Merrill C (1979) Production of a tissue-like structure by contraction of collagen lattices by human fibroblasts of different proliferative potential in vitro. Proc Natl Acad Sci U S A 76(3):1274–1278. doi:10.1073/pnas.76.3.1274
Eckes B, Krieg T, Nusgens BV et al (1995) In vitro reconstituted skin as a tool for biology, pharmacology and therapy: a review. Wound Repair Regen 3(3):248–257. doi:10.1046/j.1524-475X.1995.30304.x
Kurschat P, Zigrino P, Nischt R et al (1999) Tissue inhibitor of matrix metalloproteinase-2 regulates matrix metalloproteinase-2 activation by modulation of membrane-type 1 matrix metalloproteinase activity in high and low invasive melanoma cell lines. J Biol Chem 274(30):21056–21062. doi:10.1074/jbc.274.30.21056
Zigrino P, Mauch C, Fox JW et al (2005) Adam-9 expression and regulation in human skin melanoma and melanoma cell lines. Int J Cancer 116(6):853–859. doi:10.1002/ijc.21087
Grinnell F, Petroll WM (2010) Cell motility and mechanics in three-dimensional collagen matrices. Annu Rev Cell Dev Biol 26:335–361. doi:10.1146/annurev.cellbio.042308.113318
Eckes B, Dogic D, Colucci-Guyon E et al (1998) Impaired mechanical stability, migration and contractile capacity in vimentin-deficient fibroblasts. J Cell Sci 111(Pt 13):1897–1907
Grinnell F (1994) Fibroblasts, myofibroblasts, and wound contraction. J Cell Biol 124(4):401–404. doi:10.1083/jcb.124.4.401
Smola H, Thiekotter G, Fusenig NE (1993) Mutual induction of growth factor gene expression by epidermal-dermal cell interaction. J Cell Biol 122(2):417–429. doi:10.1083/jcb.122.2.417
Barczyk M, Carracedo S, Gullberg D (2010) Integrins. Cell Tissue Res 339(1):269–280. doi:10.1007/s00441-009-0834-6
Zhang ZG, Bothe I, Hirche F et al (2006) Interactions of primary fibroblasts and keratinocytes with extracellular matrix proteins: contribution of alpha2beta1 integrin. J Cell Sci 119(Pt 9):1886–1895. doi:10.1242/jcs.02921
Zweers MC, Davidson JM, Pozzi A et al (2007) Integrin alpha2beta1 is required for regulation of murine wound angiogenesis but is dispensable for reepithelialization. J Invest Dermatol 127(2):467–478. doi:10.1038/sj.jid.5700546
Popova SN, Barczyk M, Tiger CF et al (2007) Alpha11 beta1 integrin-dependent regulation of periodontal ligament function in the erupting mouse incisor. Mol Cell Biol 27(12):4306–4316. doi:10.1128/MCB.00041-07
Rosenfeldt H, Grinnell F (2000) Fibroblast quiescence and the disruption of ERK signaling in mechanically unloaded collagen matrices. J Biol Chem 275(5):3088–3092. doi:10.1074/jbc.275.5.3088
Mauch C, Hatamochi A, Scharffetter K et al (1988) Regulation of collagen synthesis in fibroblasts within a three-dimensional collagen gel. Exp Cell Res 178(2):493–503. doi:10.1016/0014-4827(88)90417-X
Mauch C, Adelmann-Grill B, Hatamochi A et al (1989) Collagenase gene expression in fibroblasts is regulated by a three-dimensional contact with collagen. FEBS Lett 250(2):301–305. doi:10.1016/0014-5793(89)80743-4
Zigrino P, Drescher C, Mauch C (2001) Collagen-induced proMMP-2 activation by MT1-MMP in human dermal fibroblasts and the possible role of alpha2beta1 integrins. Eur J Cell Biol 80(1):68–77. doi:10.1078/0171-9335-00134
Grinnell F (2003) Fibroblast biology in three-dimensional collagen matrices. Trends Cell Biol 13(5):264–269. doi:10.1016/S0962-8924(03)00057-6
Chiquet M, Matthisson M, Koch M et al (1996) Regulation of extracellular matrix synthesis by mechanical stress. Biochem Cell Biol 74(6):737–744. doi:10.1006/excr.1998.4363
Kessler D, Dethlefsen S, Haase I et al (2001) Fibroblasts in mechanically stressed collagen lattices assume a “synthetic” phenotype. J Biol Chem 276(39):36575–36585. doi:10.1074/jbc.M101602200
Trachslin J, Koch M, Chiquet M (1999) Rapid and reversible regulation of collagen XII expression by changes in tensile stress. Exp Cell Res 247(2):320–328. doi:10.1006/excr.1998.4363
Tomasek JJ, Gabbiani G, Hinz B et al (2002) Myofibroblasts and mechano-regulation of connective tissue remodelling. Nat Rev Mol Cell Biol 3(5):349–363. doi:10.1038/nrm809
Murad S, Grove D, Lindberg KA et al (1981) Regulation of collagen synthesis by ascorbic acid. Proc Natl Acad Sci U S A 78(5):2879–2882
Rittié L, Fisher GJ (2005) Isolation and culture of skin fibroblasts. Methods Mol Med 117:83–98. doi:10.1385/1-59259-940-0:083
Kuroda Y, Wakao S, Kitada M et al (2013) Isolation, culture and evaluation of multilineage-differentiating stress-enduring (muse) cells. Nat Protoc 8(7):1391–1415. doi:10.1038/nprot.2013.076
Acknowledgments
We thank all the members of the dermatology department in Cologne for their continued stimulating discussion; in particular we are grateful to Cornelia Mauch and Thomas Krieg for their constructive criticism and support. Work in the Eckes and Zigrino labs is supported by the German Research Foundation (Deutsche Forschungsgemeinschaft) through SFB 829.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Eckes, B., Wang, F., Rittié, L., Scherr, G., Zigrino, P. (2017). Cell-Populated Collagen Lattice Models. In: Rittié, L. (eds) Fibrosis. Methods in Molecular Biology, vol 1627. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7113-8_15
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7113-8_15
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7112-1
Online ISBN: 978-1-4939-7113-8
eBook Packages: Springer Protocols