Enhanced Survival of Neural Cells Embedded in Hydrogels Composed of Collagen and Laminin-Derived Cell Adhesive Peptide
Enhanced Survival of Neural Cells Embedded in Hydrogels Composed of Collagen and Laminin-Derived Cell Adhesive Peptide
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DOI：
10.1021/bc9000068
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发表时间：
2009-05-01
影响因子：
4.7
通讯作者：
Iwata, Hiroo
中科院分区：
文献类型：
--
作者：
Hiraoka, Makiko;Kato, Koichi;Iwata, Hiroo
为了开发可提高移植到中枢神经组织的神经细胞存活率的生物材料，制备了 I 型胶原蛋白水凝胶作为细胞载体。水凝胶用层粘连蛋白衍生肽进行修饰，已知该肽对 alpha3beta 整联蛋白具有亲和力，可在嵌入细胞中转导抗凋亡信号。为了修饰胶原蛋白，通过重组 DNA 技术将肽融合到胶原蛋白结合多肽结构域的 N 端或 C 端，或两个末端。通过聚丙烯酰胺凝胶电泳和圆二色光谱对嵌合蛋白进行了表征，同时通过生理条件下的表面等离振子共振分析验证了嵌合蛋白与胶原包被基质的结合。细胞培养测定表明，通过以肽密度依赖性方式掺入嵌合蛋白，显着促进神经球形成细胞与胶原包被的聚苯乙烯表面的粘附。对在水凝胶中培养 24 或 48 小时的细胞进行的活/死测定表明，肽的掺入提高了嵌入胶原水凝胶中的细胞的存活率。这些结果表明，含有层粘连蛋白衍生肽的胶原水凝胶提供了适合神经细胞生存的微环境。
To develop biomaterials that serve to improve the survival of neural cells transplanted into central nervous tissues, type I collagen-based hydrogels were prepared as a cell carrier. The hydrogels were modified with a laminin-derived peptide that is known to have an affinity for alpha3betal integrin, to transduce antiapoptotic signaling in embedded cells. For the modification of collagen, the peptide was fused to the N- or C-terminus, or both termini of a collagen-binding polypeptide domain by means of recombinant DNA technology. The chimeric proteins were characterized by polyacrylamide gel electrophoresis and circular dichroism spectroscopy, while binding of chimeric proteins to collagen-coated substrates was verified by surface plasmon resonance analysis under physiological conditions. Cell culture assays revealed that the adhesion of neurosphere-forming cells to collagen-coated polystyrene surfaces was significantly promoted by the incorporation of the chimeric proteins in a peptide-density dependent manner. The live/dead assays for cells cultured for 24 or 48 h in the hydrogels revealed that peptide incorporation improved the survival of cells embedded in collagen hydrogels. These results suggest that collagen hydrogel containing the laminin-derived peptide provides microenvironments suitable for the survival of neural cells.