Control of Chondrogenesis in Limb-Bud Cell Cultures by Bromodeoxyuridine

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Abstract
Initial exposure of cultured limb-bud cells (stage 23-24) to 5-bromo-2'-deoxyuridine (BrdU) irreversibly inhibits differentiation to cartilage under three different culture conditions. The inhibition of chondroitin sulfate synthesis is partially reversed by D-xylose in limb-bud cells after treatment with BrdU. The activities of four enzymes involved in chondroitin sulfate production were reduced in BrdU-treated cultures, but the magnitude of decrease was far less than the decrease in glycosaminoglycan synthesis. The slight increase in the turnover rate of sulfated glycosaminoglycans in BrdU-treated mesenchyme was not sufficient to account for the marked decrease in chondroitin sulfate content. The results suggest that BrdU treatment interferes with normal synthesis of chondroitin sulfate core protein in cultured limb-bud cells, but does not greatly diminish enzyme activities or UDP-sugar levels necessary for production of polysaccharide chains.