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固定化血清素:一种用于细胞培养的新型底物。

Immobilized serotonin: a novel substrate for cell culture.

作者信息

Hannan G N, McAuslan B R

出版信息

Exp Cell Res. 1987 Jul;171(1):153-63. doi: 10.1016/0014-4827(87)90258-8.

Abstract

Baby hamster kidney cells, bovine aortal endothelial cells, bovine smooth muscle cells, and chick embryo fibroblasts were all observed to attach and grow on serotonin which had been immobilized by covalent coupling to agarose beads. While growth and morphology of cells on immobilized serotonin appeared normal, a change in cell function may have occurred since the pattern of polypeptides expressed by these cells was different from that of cells grown on two other substrates: immobilized fibronectin and tissue culture plastic. By changing the composition of the fetal calf serum proteins in the growth medium it was shown that cells attach directly to immobilized fibronectin without mediation by medium components. In contrast, cells were found not to attach directly to immobilized serotonin but to attach indirectly via factors absorbed onto immobilized serotonin from fetal calf serum. The major component of this cell attachment activity was shown not to be fibronectin and was identified following separation by SDS-PAGE, electroblotting, and cell binding on nitrocellulose filters. The cell attachment activity compromises a major protein species of Mr 70,000 which is the molecular size of the recently identified serum spreading factor also called vitronectin.

摘要

仓鼠肾细胞、牛主动脉内皮细胞、牛平滑肌细胞和鸡胚成纤维细胞均被观察到能附着并生长在通过共价偶联固定于琼脂糖珠上的血清素上。虽然固定化血清素上的细胞生长和形态看起来正常,但这些细胞表达的多肽模式与在另外两种底物(固定化纤连蛋白和组织培养塑料)上生长的细胞不同,因此细胞功能可能已经发生了变化。通过改变生长培养基中胎牛血清蛋白的成分,结果表明细胞可直接附着于固定化纤连蛋白,无需培养基成分的介导。相反,发现细胞不会直接附着于固定化血清素,而是通过从胎牛血清中吸附到固定化血清素上的因子间接附着。这种细胞附着活性的主要成分不是纤连蛋白,在通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS - PAGE)分离、电印迹以及在硝酸纤维素滤膜上进行细胞结合后得以鉴定。细胞附着活性包含一种主要蛋白质,其分子量为70,000,这也是最近鉴定出的血清铺展因子(也称为玻连蛋白)的分子大小。

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