Hayman M W, Smith K H, Cameron N R, Przyborski S A
School of Biological and Biomedical Science, University of Durham, South Road, Durham DH1 3LE, UK.
J Biochem Biophys Methods. 2005 Mar 31;62(3):231-40. doi: 10.1016/j.jbbm.2004.12.001. Epub 2004 Dec 30.
Understanding neural differentiation and the development of complex neurite networks in three-dimensional matrices is critical for neural tissue engineering in vitro. In this study we describe for the first time the growth of human stem cell-derived neurons on solid polystyrene matrices coated with bioactive molecules. Highly porous foams were prepared from poly(styrene/divinylbenzene) using a high internal phase emulsion (HIPE) as a template to create the porous structure. The resulting polyHIPE matrices were readily coated with aqueous-based solutions including poly-d-lysine and laminin. Human neurons adhered well to poly-d-lysine coated surfaces and extended neural processes, however, neurite outgrowth was particularly enhanced when polymers also received a coating of laminin. These data clearly demonstrate the potential use of solid polystyrene scaffolds to create three-dimensional environments for cell growth and differentiation. We propose that these robust and stable matrices can be conveniently and routinely used in the tissue culture laboratory to study the behaviour of cells grown in three-dimensions.
了解神经分化以及三维基质中复杂神经突网络的发育对于体外神经组织工程至关重要。在本研究中,我们首次描述了人类干细胞衍生的神经元在涂有生物活性分子的固体聚苯乙烯基质上的生长情况。使用高内相乳液(HIPE)作为模板由聚(苯乙烯/二乙烯基苯)制备高度多孔的泡沫,以创建多孔结构。所得的聚HIPE基质很容易被包括聚-D-赖氨酸和层粘连蛋白在内的水基溶液包被。人类神经元很好地粘附在聚-D-赖氨酸包被的表面并延伸出神经突起,然而,当聚合物也接受层粘连蛋白包被时,神经突生长特别增强。这些数据清楚地证明了固体聚苯乙烯支架在为细胞生长和分化创造三维环境方面的潜在用途。我们建议这些坚固且稳定的基质可方便且常规地用于组织培养实验室,以研究在三维空间中生长的细胞的行为。