Fibronectin production by cultured human lung fibroblasts in three-dimensional collagen gel culture

Abstract

In vivo, fibroblasts are distributed in a three-dimensional (3-D) connective tissue matrix. Fibronectin is a major product of fibroblasts in routine cell culture and is thought to regulate many aspects of fibroblast biology. In this context, we sought to determine if the interaction of fibroblasts with a 3-D matrix might affect fibronectin production. To examine this hypothesis, fibronectin production by fibroblasts cultured in a 3-D collagen gel or on plastic dishes was measured by ELISA. Fibroblasts in 3-D gel culture produced more fibronectin than those in monolayer culture. Fibroblasts in 3-D culture produced increasing amounts of fibronectin when the collagen concentration of the gel was increased. The 3-D nature of the matrix appeared to be crucial because plating the fibroblasts on the surface of a plastic dish underneath a collagen gel was not different from plating them on a plastic dish in the absence of collagen. In addition to increased fibronectin production, the distribution of the fibronectin produced in 3-D culture was different from that of monolayer culture. In monolayer culture, more than half of the fibronectin was released into the culture medium. In 3-D culture, however, approximately two-thirds remained in the collagen gel. In summary, the presence of a 3-D collagen matrix increases fibroblast fibronectin production and results in greater retention of fibronectin in the vicinity of the producing cells.

Prostacyclin Analogs Inhibit Fibroblast Contraction of Collagen Gels through the cAMP-PKA Pathway

“…Only the fibronectin released into the culture media was measured in the current study. Previous studies have demonstrated that approximately one-third of fibronectin is released into the culture medium in three-dimensional culture collagen gel, and more than half is released into the culture media in monolayer culture (30). Thus the absorption amounts measured do not reflect the total fibronectin production.…”

Immunoprofile of Kuttner Tumor (Chronic Sclerosing Sialadenitis)

“…The interactions between fibroblasts, their secreted products, and the surrounding matrix are important in tissue remodeling for the development of scar formation and fibrosis. 15 It could be a response to injury of the gland. Type III collagen also showed an expression similar to that of fibronectin.…”

Hydrogel scaffolds asin vitromodels to study fibroblast activation in wound healing and disease

“…For the culture of fibroblasts in vitro , hydrogels have been formed from naturally-derived native ECM components, such as collagen, fibrin, and hyaluronic acid, 8 or synthetic materials functionalized with bioactive moieties, such as polyacrylamide modified with whole ECM proteins 9 or poly(ethylene glycol) (PEG) modified with protein mimetic peptides. 10 As will be elaborated below (Section B), fibroblasts traditionally have been cultured in fibrin 11,12 and type I collagen (collagen I) 13,14 to mimic the matrix present during different phases of wound healing. In these models, cell proliferation, activation, matrix synthesis, and contractility have been examined as signs of fibroblast activation.…”