Defective adhesion to extracellular matrix leads to altered social behaviour in cultured fibroblasts

Abstract

We describe the properties of variant mouse fibroblasts selected for poor adhesion to growth substratum containing subcellular matrix accumulated by adherent cells at confluence. The variant cells adhere to virgin plastic and grow normally to confluence in the presence of serum. After subculture and reseeding onto the same surface the cells initially adhere, but after a further 2 days of growth they retract into aggregates and detach. If the aggregates are dispersed and cells reseeded onto the same surface they remain rounded. However, if the same cells are added back to virgin plastic they adhere and grow normally. The retraction can be abolished by treating the subcellular matrix-coated plastic with papain. This behaviour therefore reflects the ability of the cells to modify the composition of the underlying substratum during growth. The variant cells also exhibit retraction 2 days after seeding on a surface previously containing wild-type cells at confluence, while wild-type cells do not retract on subcellular matrix deposited by variants. This shows that the variant behaviour arises not from a deficiency in the subcellular matrix, but from an alteration in the adhesive capacity of the cells. The results are interpreted in terms of three putative adhesion mechanisms: cell-cell adhesion in confluent monolayers and aggregates; ‘early’ type cell-substratum adhesion, which occurs during culture on virgin plastic; and ‘late’ type cell-substratum adhesion occurring on surfaces containing accumulated subcellular matrix. The variant phenotype is characterized by a deficiency in the last of these. It is also associated with an increased ability to grow in suspension culture at high dilution.