Identification of peptide sequences and protein domains derived from the glycoprotein tenascin-C for the fabrication of bioactive hydrogels to study neural stem/progenitor cell behavior in 2D and 3D cell experiments.

在我们以前的工作中，我们已经证明细胞外基质TNC(tenascin-C)的糖蛋白调节发育中的胚胎脊髓中NSPC的细胞周期长度。我们有初步证据表明，成年NSPC在纯化的TNC底物上表现不同，并提出神经干细胞群体对其ECM生态位微环境的反应是不同的。基于我们的结果，我们提出了一个研究TNC结构特征的项目，旨在识别影响NSPC维持和分化的衍生结构域和肽序列基序。在成功鉴定了蛋白质结构域和多肽序列后，它们将被用于制备具有明确组成和性质的人工水凝胶。在二维和三维细胞实验中，我们的目标是评估这些多肽和蛋白质结构域如何影响NSPC的行为。在这种情况下，结合其他水凝胶特性，如硬度、电荷和其他多肽的存在将是非常有意义的，因为这些特性可以协同作用于NSPC的行为。该研究提案的目的是确定一组最佳的水凝胶参数和功能化，以允许控制NSPC的维护和差异化。

In our previous work we have shown that the glycoprotein of the extracellular matrix tenascin-C (Tnc) modulates the cell cycle length of NSPCs in the developing embryonic spinal cord. We have preliminary evidence that adult NSPCs behave differently on purified Tnc-substrates and propose that neural stem cell populations are heterogeneous with regard to their response towards their ECM niche microenvironments. Based on our results, we submit a project that addresses the structural features of Tnc and aims at the identification of derived domains and peptide sequence motifs that affect NSPC maintenance and differentiation. After the successful identification of protein domains and peptide sequences they will be used to fabricate artificial hydrogels with well-defined composition and properties. In 2-dimensional and 3-dimensioanl cell experiments we aim to assess how these peptides and protein domains affect the behaviour of NSPCs. In this context, the combination with other hydrogel characteristics such as stiffness, charge and the presence of further peptides will be of great interest since these properties can act synergistically on NSPC behaviour. The aim of the research proposal is to identify an optimal set of hydrogel parameters and functionalisation that is allows to control NSPC maintenance and differentiation.