Cell adhesion ability of artificial extracellular matrix proteins containing a long repetitive Arg-Gly-Asp sequence
Cell adhesion ability of artificial extracellular matrix proteins containing a long repetitive Arg-Gly-Asp sequence
复制标题
DOI：
10.1263/jbb.100.82
复制
发表时间：
2005-07-01
影响因子：
2.8
通讯作者：
Nagamune, T
中科院分区：
文献类型：
--
作者：
Kurihara, H;Nagamune, T
我们产生了含有重复Arg-Gly-Asp（RGD）序列的重组人工细胞外基质（ECM）蛋白：双（RGD 2）、21（RGD 21）和43（RGD 43）重复的RGD。RGD 43包被的载玻片促进成纤维细胞NIH 3 T3细胞在表面上的粘附和铺展。由于在粘附在RGD 43包被的载玻片上的细胞中也观察到肌动蛋白应力纤维和焦点接触，因此认为RGD 43中的RGD肽通过整合素传递粘附信号并促进细胞粘附。我们包被重组ECM蛋白，每个含有重复的RGD结构域，在聚苯乙烯板上，并使用三种不同的细胞系，即成纤维细胞NIH 3 T3，HeLa癌和神经元PC 12细胞系，研究RGD长度对细胞粘附能力的影响。结果表明，RGD 43的细胞粘附能力上级优于天然细胞外基质蛋白、纤连蛋白和层粘连蛋白，尽管RGD重复序列长度对细胞粘附能力的影响取决于细胞系。作为用于细胞培养的人工三维支架，我们还通过使用戊二醛的交联反应制备了RGD 43水凝胶。在RGD 43水凝胶支架上，成纤维细胞也在无血清条件下成功粘附。
We generated recombinant artificial extracellular matrix (ECM) proteins containing repetitive Arg-Gly-Asp (RGD) sequences: double (RGD2), 21 (RGD21) and 43 (RGD43) repeats of RGD. RGD43-coated glass slides promoted fibroblast NIH3T3 cell adhesion and spreading on the surface. Since actin stress fibers and focal contacts were also observed in cells adhering on RGD43-coated glass slides, it was suggested that the RGD peptides in RGD43 transmitted an adhesion signal via integrins and promoted cell adhesion. We coated recombinant ECM proteins, each containing repetitive RGD domains, on polystyrene plates and investigated the effects of RGD length on the cell adhesion ability using three different cell lines, namely, fibroblast NIH3T3, HeLa cancer and neuronal PC12 cell lines. The results indicated that RGD43 had a cell adhesion ability superior to those of natural extracellular matrix proteins, fibronectin and laminin, although the effects of RGD repeat length on the cell adhesion ability depended on the cell line. As an artificial three-dimensional scaffold for cell cultivation, we also prepared an RGD43 hydrogel by a crosslinking reaction using glutaraldehyde. On the RGD43 hydrogel scaffold, fibroblast cells also successfully adhered under serum-free conditions.