Development of a new serum-free medium, USC-HC1, for growth and normal phenotype in postembryonic chicken growth plate chondrocytes.
Development of a new serum-free medium, USC-HC1, for growth and normal phenotype in postembryonic chicken growth plate chondrocytes.
复制标题
开发一种新的无血清培养基 USC-HC1，用于胚胎后鸡生长板软骨细胞的生长和正常表型。
DOI：
10.1007/bf02623519
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Wuthier,RE
中科院分区：
文献类型：
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作者：
Hale,LV;Hale,JE;Kemick,ML;Ishikawa,Y;Wuthier,RE
以104 MCDB培养基为基础，研制出鸡胚胎后骨骺生长板软骨细胞的无血清培养基。为了使这些挑剔的细胞能够存活、生长并表达正常表型，许多氨基酸的水平需要比 MCDB 104 大幅提高，并且需要改变缓冲系统并添加 SerXtend（一种含有多种生长因子（包括成纤维细胞生长因子）的成分明确的无血清产品）。还需要提供细胞附着因子，可以通过用 II 型胶原蛋白涂覆培养物表面，或者更好的是，让新鲜释放的细胞在铺板前在补充有 10% 胎牛血清的培养基中恢复几个小时。这种新型无血清培养基，我们称之为 USC-HC1，支持软骨细胞的生长和复制、正常多边形形态的保留、细胞碱性磷酸酶活性的显着水平表达、硫酸化蛋白聚糖、II 型胶原的产生以及富含碱性磷酸酶的基质囊泡的形成。然而，USC-HC1 的主要优点是，它将首次提供一个机会，在胎牛血清中不存在可变（刺激和抑制）因子的情况下，检查各种确定的生长和激素因子对生长板软骨细胞表型表达和分化的影响。
A serum-free medium for postembryonic chicken epiphyseal growth plate chondrocytes has been developed from 104 MCDB medium. To enable these fastidious cells to survive, grow, and express normal phenotype, a substantial increase over MCDB 104 in the level of many of the amino acids was required, as well as a change in the buffer system and the addition of SerXtend, a defined, serum-free product containing various growth factors, including fibroblast growth factor. Also required was the provision of cell attachment factors, either by coating culture surfaces with type II collagen, or better, by allowing the freshly released cells to recover for several hours in a medium supplemented with 10% fetal bovine serum before plating. Ths new serum-free medium, which we call USC-HC1, supports growth and replication, the retention of normal polygonal morphology, the expression of significant levels of cellular alkaline phosphatase activity, the production of sulfated proteoglycans, type II collagen, and the formation of alkaline phosphatase-rich matrix vesicles by the chondrocytes. The major advantage of USC-HC1, however, is that it will provide for the first time an opportunity to examine the effects of various defined growth and hormonal factors on the phenotypic expression and differentiation of growth plate chondrocytes, in the absence of the variable (stimulatory and inhibitory) factors present in fetal bovine serum.