Summary
We have developed a procedure to explant fibroblasts from the anterior cruciate ligament (ACL) and the medial collateral ligament (MCL) of the rabbit knee, and have optimized conditions for maintaining them in culture. Maximal growth for both ACL and MCL cells was obtained with Dulbecco's modified Eagle's medium supplemented with 15% fetal bovine serum and 250 μM ascorbate. ACL and MCL fibroblasts displayed intrinsic differences in their responses to changes in culture parameters. Specifically, they displayed different growth responses when plated at different densities and responded to RPMI 1640 medium in very different ways. There were also biochemical differences between the cell types. Both cell types produced similar amounts of collagen in culture, but the ratio of type I to type III, the major collagen subtypes produced by these cells, were different. ACL fibroblasts produced 86.7% type I and 13.3% type III, and MCL fibroblasts produced 71.1% type I and 28.9% type III. In addition, total protein produced by ACL fibroblasts was higher than that produced by MCL cells. This confirms the suggestions of previous researchers that such differences might exist.
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This work was funded by a grant-in-aid from Medtronic of Canada, by an R&D Grant from the Alberta Ministry of Technology, Research and Telecommunications, and by the Alberta Heritage Foundation for Medical Research.
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Ross, S.M., Joshi, R. & Frank, C.B. Establishment and comparison of fibroblast cell lines from the medial collateral and anterior cruciate ligaments of the rabbit. In Vitro Cell Dev Biol 26, 579–584 (1990). https://doi.org/10.1007/BF02624206
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DOI: https://doi.org/10.1007/BF02624206