Sulfate incorporation by articular chondrocytes in monolayer culture.
Sulfate incorporation by articular chondrocytes in monolayer culture.
复制标题
单层培养中关节软骨细胞掺入硫酸盐。
作者：
L. Sokoloff;C. Malemud;W. T. Green
原代单层或传代培养的关节软骨细胞合成硫酸盐化的大分子，推测是粘多糖。生后兔软骨细胞内~(35)SO4掺入在孵育24小时时达到稳定水平；细胞外不可透析性(离心液+胰酶消化)计数在72小时后增加，达到细胞内值的100倍。细胞间比较，软骨细胞各组分的放射性硫酸盐计数均高于成纤维细胞。来自一只3岁大的兔子的数据与最近断奶的动物的数据相当。在成人和牛关节软骨细胞的培养中也发现了硫酸盐化粘多糖的分泌。尽管Ham的F12培养液有利于建立软骨细胞的原代培养体系，但在继代培养中使用Dulbecco-Eagle培养基加或不加L-脯氨酸时，硫酸盐掺入率更高。在已经融合的单层中加入抗坏血酸(5 mg/100毫升)没有一致的效果。
Articular chondrocytes in primary monolayers or subcultures synthesize sulfated macromolecules, presumably mucopolysaccharides. Intracellular 35SO4 incorporation by postnatal rabbit chondrocytes reached a steady level by 24 hr of incubation; the nondialyzable extracellular (centrifuged medium + trypsin digest) counts increased over 72 hr, when they were up to 100 times the intracellular value. Cell for cell, radiosulfate counts in each fraction were higher in chondrocytes than fibroblasts. Data from a 3-year-old rabbit were comparable to those from recently weaned animals. Secretion of sulfated mucopolysaccharide was also found in cultures of adult human and bovine articular chondrocytes. Although Ham's F12 medium is advantageous in establishing a primary culture of chondrocytes, sulfate incorporation was greater when Dulbecco-Eagle medium, with or without supplemental L-proline, was used for subcultures. Ascorbic acid (5 mg/100 ml) added to an already confluent monolayer had no consistent effect.