Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, The Biomedical Centre, S-75123 Uppsala, Sweden.
Biomaterials. 2013 Nov;34(33):8223-34. doi: 10.1016/j.biomaterials.2013.07.058. Epub 2013 Aug 2.
Silk matrices have previously been shown to possess general properties governing cell viability. However, many cell types also require specific adhesion sites for successful in vitro culture. Herein, we have shown that cell binding motifs can be genetically fused to a partial spider silk protein, 4RepCT, without affecting its ability to self-assemble into stable matrices directly in a physiological-like buffer. The incorporated motifs were exposed in the formed matrices, and available for binding of integrins. Four different human primary cell types; fibroblasts, keratinocytes, endothelial cells and Schwann cells, were applied to the matrices and investigated under serum-free culture conditions. Silk matrices with cell binding motifs, especially RGD, were shown to promote early adherence of cells, which formed stress fibers and distinct focal adhesion points. Schwann cells acquired most spread-out morphology on silk matrices with IKVAV, where significantly more viable cells were found, also when compared to wells coated with laminin. This strategy is thus suitable for development of matrices that allow screening of various cell binding motifs and their effect on different cell types.
丝基质先前已被证明具有普遍的细胞活力调控特性。然而，许多细胞类型也需要特定的黏附位点才能成功进行体外培养。在此，我们表明可以将细胞结合基序通过基因融合到部分蜘蛛丝蛋白 4RepCT 上，而不会影响其在类似于生理的缓冲液中自我组装成稳定基质的能力。所融合的基序在形成的基质中暴露，可用于整合素的结合。四种不同的人原代细胞类型；成纤维细胞、角质形成细胞、内皮细胞和许旺细胞，被应用于基质，并在无血清培养条件下进行了研究。含有细胞结合基序（尤其是 RGD）的丝基质可促进细胞的早期黏附，细胞形成应力纤维和明显的黏附斑。许旺细胞在含有 IKVAV 的丝基质上获得了最伸展的形态，在这些基质上发现了更多有活力的细胞，与铺有层粘连蛋白的孔相比也是如此。因此，该策略适用于开发允许筛选各种细胞结合基序及其对不同细胞类型影响的基质。
