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

基于DNA微阵列技术的polySia基质对细胞系统影响的交互式测试和优化

• 批准号: 5432453
• 负责人: Professor Dr. Thomas Scheper
• 金额: --
• 依托单位: Institut für Neuroanatomie und Zellbiologie
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2004
• 资助国家: 德国
• 起止时间: 2003-12-31 至 2007-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5432453?language=en
• 关键词: Interacitve; testing; optimization; polySia; matrices

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.

众所周知，细胞以改变的基因表达响应于变化的环境，并且修饰的polySia基质可以影响培养细胞的增殖和分化。在本项目中，将为WP 1和WP 2中产生的可溶性天然和非天然polySia及其生成的基质（WP 2和WP 3）建立细胞（以及随后的动物）试验系统。材料将测试之前或之后定义的装饰与生长因子，细胞因子，各种特定的发射机，和其他。这些基质与细胞系统的相互作用和影响将被监测，特别是关于使用“定制”微阵列的基因表达。这些特异性微阵列将允许同时分析数百种基因的表达，包括神经特异性蛋白质如神经丝、受体或神经递质相关酶、细胞外基质、附着和粘附蛋白、细胞因子及其受体，以及一组对照基因。在对生长和分化细胞的标准表达模式进行评估后，这些模式可用于控制不同培养条件下的细胞行为。这种分析允许非常详细地了解细胞的状态，从而允许将细胞生物学和基质质量相关联。根据获得的数据，应能够对polySia矩阵进行迭代优化。此外，将分别使用生理神经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假定的细胞来源。除新生和成年大鼠细胞外，还将评价polySia基质和支架对成年人雪旺细胞的影响。