D'Arville C, Mason R M
Biochim Biophys Acta. 1983 Oct 4;760(1):53-60. doi: 10.1016/0304-4165(83)90123-x.
Incorporation of [3H]glucosamine into hyaluronate synthesized by chondrocyte cultures was dependent on the concentration of foetal calf serum in the culture medium. [3H]Hyaluronate levels in cultures supplemented with 2% serum, or maintained without serum, were about 60 and 43%, respectively, of that in cultures maintained with 15% serum. Addition of insulin to cultures maintained with 15% serum had no significant effect on [3H]hyaluronate synthesis. Addition of the hormone to cultures maintained with 2% serum increased [3H]hyaluronate synthesis to levels either the same (1 ng insulin/ml), or greater than (100 ng insulin/ml) that in cultures maintained with 15% serum. The [3H]hyaluronate synthesized by the cultures was of very high molecular weight irrespective of the level of synthesis. [3H]Hyaluronate formed about 12% of the total [3H]glycosaminoglycan synthesized under all culture conditions. Synthesis of 35S, 3H-labelled proteoglycan was reduced, or increased, by the same relative amounts as [3H]hyaluronate, under the different culture conditions. Incorporation of [3H]glucosamine into hyaluronate by near confluent cultures of fibroblasts derived from the Swarm rat chondrosarcoma was reduced by 50% in cultures treated with 2% foetal calf serum compared to those maintained with 15% serum. [3H]Hyaluronate synthesis by fibroblast cultures treated with 2% serum was not stimulated by addition of insulin.
软骨细胞培养合成的透明质酸中[3H]葡糖胺的掺入量取决于培养基中胎牛血清的浓度。补充2%血清的培养物或无血清培养的[3H]透明质酸水平分别约为补充15%血清培养物的60%和43%。向补充15%血清的培养物中添加胰岛素对[3H]透明质酸的合成没有显著影响。向补充2%血清的培养物中添加该激素可使[3H]透明质酸的合成增加至与补充15%血清的培养物相同（胰岛素浓度为1 ng/ml）或更高（胰岛素浓度为100 ng/ml）的水平。无论合成水平如何，培养物合成的[3H]透明质酸分子量都非常高。在所有培养条件下，[3H]透明质酸约占合成的总[3H]糖胺聚糖的12%。在不同培养条件下，35S、3H标记的蛋白聚糖的合成减少或增加的相对量与[3H]透明质酸相同。与补充15%血清的培养物相比，用2%胎牛血清处理的来自Swarm大鼠软骨肉瘤的成纤维细胞近汇合培养物中，[3H]葡糖胺掺入透明质酸的量减少了50%。用2%血清处理的成纤维细胞培养物中添加胰岛素不会刺激[3H]透明质酸的合成。
