Interacitve testing and optimization of polySia matrices effects on cell systems based on DNA microarray techniques

众所周知，细胞通过改变基因表达来响应不断变化的环境，修饰的PolySia基质可以影响培养细胞的增殖和分化。在该项目中，将为WP1和WP2中生产的可溶性天然和非天然聚SIA及其生成的基质(WP2和WP3)建立细胞(以及稍后的时间点也包括动物)测试系统。材料将在限定的装饰之前或之后进行测试，包括生长因子、细胞因子、各种特定的传递体等。将监测这些基质与细胞系统的相互作用和影响，特别是关于基因表达的情况，使用“量身定制”的微阵列。这些特定的微阵列将允许同时分析数百个基因的表达，包括神经特异蛋白，如神经丝、受体或神经递质相关酶、细胞外基质、附着和黏附蛋白、细胞因子及其受体，以及一组控制基因。在对生长和分化细胞的标准表达模式进行评估后，这些模式可以用于控制细胞在不同培养条件下的行为。这种分析可以非常详细地洞察细胞的状态，因此可以将细胞生物学和基质质量联系起来。基于所获得的数据，应该能够对PolySia矩阵进行迭代优化。此外，将分别使用生理性神经细胞和神经胶质细胞对基质和支架进行生物学评价，这为神经损伤后的细胞替代提供了可能的细胞来源。除了新生和成年大鼠细胞外，还将评估PolySia基质和支架对成年人类雪旺细胞的影响。

It is well known that cells respond with altered gene expression to changing environments and that modified polySia matrices can influence the proliferation and differentiation of cultured cells. Within this project cellular (and to a later time point also animal) test systems will be set up for soluble natural and non-natural polySia produced in WP1 and WP2 and matrices generated thereof (WP2 and WP3). Materials will be tested before or after defined decoration with growth factors, cytokines, various specific transmitters, and others. The interaction and influence of these matrices with the cell systems will be monitored especially with regard to gene expression using "tailor-made" microarrays. These specific microarrays, will allow the simultaneous analysis of the expression of several 100 genes including neural specific proteins like neurofilaments, receptors or neurotransmitter related enzymes, extracellular matrix, attachment and adhesion proteins, cytokines and their receptors, as well as a panel of control genes. After standard expression patterns have been evaluated for growing and differentiating cells, these patterns can be used to control cell behavior under variant culture conditions. This kind of analysis allows a very detailed insight into the status of a cell and thus allows correlating cell biology and quality of matrices. Based on the obtained data the iterative optimization of polySia matrices shall be made possible. Furthermore, biological evaluation of matrices and scaffolds will be performed using physiological nerve and glial cells, respectively, which provide putative cell sources for cell substitution after nerve lesion. In addition to neonatal and adult rat cells, the effects of polySia matrices and scaffolds will be also evaluated on adult human Schwann cells.