Type IV Collagen and Fibronectin Enhance Human Keratinocyte Thymidine Incorporation and Spreading in the Absence of Soluble Growth Factors

In various cell culture systems, extracellular matrix components have been demonstrated to be mitogenic and, in some cases, to substitute for growth factors. In order to study the effects of various matrices on keratinocyte growth, we assessed the incorporation of tritiated thymidine and cell number on short-term cultures of human keratinocytes plated on different substrata. For determination of whether thymidine incorporation by keratinocytes was related to the ability of the cells to attach and spread on the substratum, experiments to determine the percentage of attached and spread cells on each matrix surface were performed. High levels of attachment and incorporation of thymidine with no preferential attachment to a given matrix were evident when the cells were cultured in the presence of growth factors. When growth factors were absent, keratinocytes likewise showed no preferential attachment to a given matrix component, but demonstrated enhanced thymidine incorporation when apposed to type IV collagen or fibronectin in comparison with tissue culture plastic or laminin. In the absence of epidermal growth factor (EGF) and bovine pituitary extract (BPE), increased spreading on type IV collagen and fibronectin was associated with enhanced incorporation of thymidine. In agreement with the thymidine incorporation results, when keratinocytes were cultured for 7 d, cell numbers were increased in cultures plated on type IV collagen only if growth factors were excluded from the medium. When attachment of cells to substrata with or without growth factors was compared, either EGF or BPE enhanced attachment to all of the substrata tested. It is concluded that under suboptimal growth conditions extracellular matrix components can modulate keratinocyte growth. Also, under these conditions, spreading, but not attachment, correlates with growth potential.