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There are several different culture models to study the ability of fibroblasts to reorganize and contract collagen matrices in vitro. In the floating contraction model, a freshly polymerized collagen matrix containing cells is released from the culture dish and allowed to float in culture medium, and contraction occursin the absence of external mechanical load and without appearance of stress fibers in the cells. In the attached model, a polymerized collagen matrix containing cells remains attached to the culture dish during contraction. Mechanical tension develops during contraction, and cellular stress fibers assemble. The two-step model combines an initial period of attached matrix contraction leading to mechanical loading, followed by release of the matrices, resulting in mechanical unloading and further contraction as mechanical stress dissipates.
3D collagen matrix has also been used in the studies of integrin signaling, cell apoptosis and cytoskeleton reorganization. Since three-dimensional matrix adhesions differ in structure, localization, and function from two-dimensional adhesions; and therefore, three-dimensional cell-matrix interactions may be more relevant biologically.
Kit Components
1. Collagen Solution (Part No. 20101): One 10 mL bottle of sterile bovine Type I Collagen at 3.0 mg/mL
2. Neutralization Solution (Part No. 20102): One 0.5 mL tube
3. 5X DMEM Medium (Part No. 20103): One 5 mL bottle
4. 5X PBS (Part No. 20104): One 5 mL bottle
5. 100X Cell Contraction Inhibitor (Part No. 20105): One 1 mL tube of 1M 2, 3-Butanedione Monoxime (BDM) in DMSO
Storage
Store all components at 4ºC until their expiration dates.
