Reconstituted type V collagen fibrils as cementing materials in the formation of cell clumps in culture

Abstract

Previous studies have reported that type V collagen is an anti-adhesive substrate for cultured cells in that the cells detach from culture dishes coated with type V collagen molecules or polypeptides derived from them. We have noticed that human fetal lung fibroblasts (TIG-1) initially show no reduction in adherence to and spreading on a dish coated with reconstituted type V collagen fibrils but eventually detach from the dish and form cell clumps. To determine the way in which reconstituted type V collagen fibrils are involved in cell clump formation, we have followed the fate of the fluorescence of type V collagen fibrils pre-labeled with fluorescein isothiocyanate. Essentially, all the fluorescence disappeared from the dish surface as the cells detached and was condensed in the cell clumps. The cells that were recovered from clumps and dissociated into separate cells by trypsin treatment proliferated normally after they were seeded on a bare culture dish. This result and those from gel electrophoresis, fluorescence microscopy, and a cell proliferation assay indicate that the cell detachment from the dish is not caused by cell necrosis or apoptosis but by cellular motility together with the unique features of type V collagen fibrils. Not only the adherence of type V collagen fibrils to TIG-1 cells is much stronger than that to the culture dish, but the fibrils are retained on the cellular surface. The strong adherence of type V collagen fibrils to cells plays a role in cementing TIG-1 cells together.

Extraction and characterization of bovine collagen Type V and its effects on cell behaviors

“…Col. V with high concentration can make L929 fall off the culture medium and gather into clusters ( Fig. 11 ), similar to the Takanori’s result [ 33 ]. With an increase in the Col. V proportion, the diameter of collagen fibers was changed, which further affected the combination of collagen with cells.…”

Collagen type V enhances matrix contraction by human periodontal ligament fibroblasts seeded in three-dimensional collagen gels

“…In response to the collagens, the cells are modulated in their proliferation and expression of matrix metalloproteinases (MMPs) (Kerkvliet et al, 2003). Of the collagens tested it was in particular collagen V that appeared to affect fibroblast behavior (Fukuda et al, 1988;Hatai et al, 1993;Kerkvliet et al, 2003;Kihara et al, 2004;Luparello et al, 1990;Ruggiero et al, 1994Ruggiero et al, , 1996Sakata et al, 1992Sakata et al, , 1994Yamamoto et al, 1992).…”

“…Most studies focusing on the effects of the different collagen types in relation to cell activity made use of monolayer cultures (Kerkvliet et al, 2003;Kihara et al, 2004;Ruggiero et al, 1994Ruggiero et al, , 1996. In order to approximate the in vivo situation, fibroblasts are seeded in a 3D collagen matrix in vitro.…”

Fragility of Reconstituted Type V Collagen Fibrils with the Chain Composition of α1(V)α2(V)α3(V) Respective of the D-Periodic Banding Pattern

“…The type Vp collagen prevents the attachment, spreading [31], and proliferation [32] of endothelial cells and of porcine vascular smooth muscle cells in vitro [33]. Kihara et al [34,35] have shown that the reconstituted fibrils from pepsin-extracted type V collagen subtype with the chain composition of [α1(V)] 2 α2(V) (Vp112) strongly adhere to the surface of cultured fibroblasts and form the clump aggregates consisting of cells and type Vp112 collagen fibrils. This effect is not shared with type I collagen, indicating that the reconstituted fibrils from the triple-helical domain of type V collagen have a different property compared to those of type I collagen.…”