A Mouse 3T6 Fibroblast Cell Culture Model for the Study of Normal and Protein-Engineered Collagen Synthesis and Deposition into the Extracellular Matrix

Abstract

Mouse 3T6 i ro asts deposited an organize co agenous extrace u ar matrix during longterm culture in the presence of ascorbic acid. The matrix produced by the cells had a similar comprising distribution of collagen types as the mouse dermal matrix, comprising predominantly type I with smaller amounts of types III aand V collagens. By day more than 70% of the collagen in the 3T6 matrix was involved in covalent crosslinkages and required pepsin digestion for extraction. Incorporation of NaB3H4 into reducible crosslinks and aldehydes directly demonstrated the involvement of the α1(I)CB6 an α2(I)CB3.5 in crosslinks. The pattern of reducible crosslinks in the in vitro 3T6 matrix was similar to tha..