Stimulatory effect of TGF‐β on anionic glycoconjugate synthesis by rat calvarial cells: Specificity, uncoupling of cell density dependence, and modulation by chondroitin sulfate

Abstract

Anchorage‐dependent cultures of a population of cells derived from the outer part of the rat calvaria demonstrated decreased net accumulation of radiolabeled chondroitin sulfate (CS) and hyaluronic acid (HA) per cell as the cell density of the cultures increased. The addition of TGF‐β1 resulted in large stimulations of the net CS, but not of the net HA, accumulating in the medium at all cell densities and an abolition of the density‐dependent effect. These effects were largely due to increases in newly synthesized CS appearing in the medium. Supplementation of the culture media with CS had complex but relatively small effects on the stimulation of the net accumulation of radiolabeled medium CS by TGF‐β1. The addition of TGF‐β1 also resulted in a biphasic effect on cell growth that depended on the plating density, but cell growth differences could not account for the marked stimulation of CS synthesis by TGF‐β1. Experiments with cycloheximide and β‐xyloside and isolation of the intact anionic glycoconjugates (AG) indicated that although synthesis of core protein was the limiting factor in CS synthesis, TGF‐β1 stimulated the synthesis of CS chain when sufficient β‐xyloside acceptor was available. The overall results suggest that, in this cell system, the action of TGF‐β1 on the synthesis of the major extracellular AGs is characterized by a relatively specific upregulation of CS proteoglycan (PG) synthesis and an uncoupling of the inhibitory effect of high cell density on CS PG synthesis. Copyright © 1993 ASBMR