Serum-free cultured keratinocytes fail to organize fibronectin matrix and possess different distribution of beta-1 integrins
Serum-free cultured keratinocytes fail to organize fibronectin matrix and possess different distribution of beta-1 integrins
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DOI：
10.1034/j.1600-0625.2001.010002080.x
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发表时间：
2001-04-01
影响因子：
3.6
通讯作者：
Dimoudis, N
中科院分区：
文献类型：
--
作者：
Altankov, G;Hecht, J;Dimoudis, N
用于培养角质形成细胞的无血清培养基配方的开发是实现大量表皮细胞用于实验和治疗研究的突破，特别是支持伤口愈合过程。然而，尚不清楚将细胞转变为高度增殖表型是否可能反映对伤口修复重要的其他细胞功能的变化，因为它们与细胞外基质成分的粘附相互作用。细胞外基质的重塑，特别是纤连蛋白的重塑，对于在伤口愈合过程中引导细胞发挥着重要作用。然而，组织这种临时纤连蛋白基质的分子机制仍不清楚。我们发现，含血清培养基中的角质形成细胞虽然数量少于成纤维细胞，但能够从基质中去除吸附的荧光标记纤连蛋白，并沿着细胞外围将其重组为纤维状模式。 3天后，分泌的纤连蛋白也组织成基质样纤维，并在细胞迁移后形成沉积在基质上的簇。相反，无血清培养的角质形成细胞无法组织预吸附的荧光标记的纤连蛋白以及分泌的纤连蛋白，尽管它们在这些条件下生长得很好。将细胞切换到含有血清的培养基中，开始从培养基中去除荧光标记的纤连蛋白，但不会以纤维状模式重组。最有可能的是，这些角质形成细胞通过表达蛋白水解活性来去除荧光标记的纤连蛋白，而不是通过 β (1) 整联蛋白的机械功能。后者在含有血清的条件下广泛分散，并倾向于在无血清培养细胞中组织成粘着斑。我们假设它们的瞬时表达和不同的亲和力状态可能对角质形成细胞迁移和基质组装机制很重要。
The development of serum free medium formulation for culturing keratinocytes was a breakthrough in achieving a high number of epidermal cells for experimental and therapeutic studies, in particular to support the wound healing process. It is not clear, however, if switching the cells to highly proliferative phenotype may reflect change in other cellular functions important for the wound repair as their adhesive interactions with the extracellular matrix components. Remodelling of the extracellular matrix, particularly of fibronectin plays an essential role for guiding the cells during wound healing. The molecular mechanisms for organization of this provisional fibronectin matrix, however, are still not clear. We found that keratinocytes in serum containing medium, although in fewer numbers than fibroblasts, were able to remove adsorbed fluorescent labelled fibronectin from the substratum and reorganize it in a fibrilar pattern along the cell periphery. After 3 days the secreted fibronectin had also been organized as matrix-like fibers and as clusters deposited on the substratum after migrating cells. In contrast, serum free cultured keratinocytes fail to organize pre-adsorbed fluorescent labelled fibronectin, as well as the secreted fibronectin, although they grow very well under these conditions. Switching the cells to serum containing medium initiates the removal of fluorescent labelled fibronectin from the substratum, however without reorganization in fibrillar pattern. Most likely, these keratinocytes remove fluorescent labelled fibronectin by the expression of proteolytic activity, rather than with the mechanical function of beta (1) integrins. The latter were diffusely dispersed in serum containing conditions and tend to organize in focal adhesions in serum free cultured cells. We assumed their transient expression and different affinity state might be important for the keratinocyte migration and matrix assembly mechanism.