Reconstituted type V collagen fibrils as cementing materials in the formation of cell clumps in culture.
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.
Kihara
Takanori
T
Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo, 153-8902, Japan.
Takemura
Yukitoshi
Y
Imamura
Yasutada
Y
Mizuno
Kazunori
K
Hayashi
Toshihiko
T
D002448:Cell Adhesion / Q000502:physiology*; D002449:Cell Aggregation / Q000502:physiology*; D002460:Cell Line / Q000502:physiology*; D002465:Cell Movement; D049109:Cell Proliferation; D024062:Collagen Type V / Q000737:chemistry / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000166:cytology* / Q000502:physiology; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D005453:Fluorescence; D006801:Humans; D008168:Lung / Q000166:cytology / Q000196:embryology; D008856:Microscopy, Fluorescence; D013194:Staining and Labeling
D024062:Collagen Type V; D016650:Fluorescein-5-isothiocyanate