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首页> 外文期刊>Acta biomaterialia >Thick collagen-based 3D matrices including growth factors to induce neurite outgrowth
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Thick collagen-based 3D matrices including growth factors to induce neurite outgrowth

机译：基于厚胶原蛋白的3D矩阵，包括诱导神经突向外生长的生长因子

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摘要

Designing synthetic microenvironments for cellular investigations is a very active area of research at the crossroads of cell biology and materials science. The present work describes the design and functionalization of a three-dimensional (3D) culture support dedicated to the study of neurite outgrowth from neural cells. It is based on a dense self-assembled collagen matrix stabilized by 100-nm-wide interconnected native fibrils without chemical crosslinking. The matrices were made suitable for cell manipulation and direct observation in confocal microscopy by anchoring them to traditional glass supports with a calibrated thickness of ～50 μm. The matrix composition can be readily adapted to specific neural cell types, notably by incorporating appropriate neurotrophic growth factors. Both PC-12 and SH-SY5Y lines respond to growth factors (nerve growth factor and brain-derived neurotrophic factor, respectively) impregnated and slowly released from the support. Significant neurite outgrowth is reported for a large proportion of cells, up to 66% for PC12 and 49% for SH-SY5Y. It is also shown that both growth factors can be chemically conjugated (EDC/NHS) throughout the matrix and yield similar proportions of cells with longer neurites (61% and 52%, respectively). Finally, neurite outgrowth was observed over several tens of microns within the 3D matrix, with both diffusing and immobilized growth factors.
机译：在细胞生物学和材料科学的十字路口，设计用于细胞研究的合成微环境是一个非常活跃的研究领域。本工作描述了致力于研究神经细胞神经突生长的三维（3D）培养支持物的设计和功能化。它基于密集的自组装胶原蛋白基质，该基质由100 nm宽的相互连接的天然原纤维稳定，无需化学交联。通过将它们固定在标定厚度约为50μm的传统玻璃支架上，使其适合于在共聚焦显微镜下进行细胞处理和直接观察。基质组成可以容易地适应特定的神经细胞类型，特别是通过掺入适当的神经营养生长因子。 PC-12和SH-SY5Y株系均对从支持物中浸渍并缓慢释放的生长因子（分别为神经生长因子和脑源性神经营养因子）产生反应。据报道，大部分细胞都有明显的神经突生长，PC12高达66％，SH-SY5Y高达49％。还显示了两种生长因子都可以在整个基质中化学偶联（EDC / NHS），并产生相似比例的具有更长神经突的细胞（分别为61％和52％）。最终，在3D矩阵内观察到了神经突长出数十微米，同时扩散和固定的生长因子。

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