In the early 1970s it was discovered that fibroblasts from human foreskins cultured in or on top of hydrated collagen gels led to gel contraction. Contraction occurs by physical rearrangement of the collagen fibrils (rather than collagen degradation) within the collagen gels by the fibroblasts1. The rearrangement is also dependent on the presence of serum which allows binding of the cells to the collagen fibrils1. The key integrins responsible for this cell-collagen interaction are the α1 and α2 chains of the β1 subfamily2.
Appropriate cells are suspended in 3 mg/ml collagen solution and rapidly pHed to neutrality with sodium hydroxide. The cell/gel mixture is pipeted into a well. After solidification, the gel is gently dissociated from the well’s walls and bottom and allowed to freely float in culture media in the well. At various times during cell growth, image wells and quantitate contraction.
Cell Types Used
heart fibroblast contraction2
smooth muscle-like cell contraction4
- Guidry C and Grinnell F, Studies on the mechanism of hydrated collagen gel reorganization by human skin fibroblasts. J. Cell Sci (1985) 79: 67-81.
- Carver W et al, Development of cardiovascular systems: molecules to organisms, Burggren WW, Keller BB (editors), Cambridge University Press 1997.
- Ngo P et al, Collagen Gel Contraction Assay, Cell-Cell Interactions: methods and protocols (S. Colgan, ed.), Humana Press, Totowa NJ 2006.
- Jeon ES et al, A Rho Kinase/Myocardin-Related Transcription Factor-A_Dependent Mechanism Underlies the Sphingosylphosphorylcholine-Induced Differentiation of Mesenchymal Stem Cells Into Contractile Smooth Muscle Cells. Circ. Res. (2008) 103: 635-642