PASTEUR, Département de chimie, École normale supérieure, UPMC Univ. Paris 06, CNRS, PSL Research University, 75005 Paris, France. Sorbonne Universités, UPMC Univ. Paris 06, École normale supérieure, CNRS, PASTEUR, 75005 Paris, France.
Biomed Mater. 2018 Feb 28;13(3):034105. doi: 10.1088/1748-605X/aaa777.
Tissue engineering strives to create functional components of organs with different cell types in vitro. One of the challenges is to fabricate scaffolds for three-dimensional (3D) cell culture under physiological conditions. Of particular interest is the investigation of the morphology and function of the central nervous system cultured using such scaffolds. Here, we used an elastomer-polydimethylsiloxane (PDMS)-to produce lattice-type scaffolds from a photolithography-defined template. The photomask with antidot arrays was spin-coated by a thick layer of resist, and was downward mounted on a rotating stage at an angle of 45°. After the exposure was repeated three or more times, maintaining the same exposure plan but rotated by the same angle, a photoresist was developed to produce a 3D porous template. Afterwards, a pre-polymer mixture of PDMS was poured in and cured, followed by a resist etch, resulting in lattice-type PDMS features. Before cell culture, the PDMS lattices were surface functionalized. A culture test was conducted using NIH-3T3 cells and primary hippocampal cells from rats, showing homogenous cell infiltration and 3D attachment. As expected, a much higher cell number was found in the 3D PDMS lattices compared to the 2D culture. We also found a higher neuron-to-astrocyte ratio and a higher degree of cell ramification in the 3D culture compared to the 2D culture due to the change of scaffold topography and the elastic properties of the PDMS micro-lattices. Our results demonstrate that the 3D PDMS micro-lattices improve the survival and growth of cells, as well as the network formation of neurons. We believe that such an enabling technology is useful for research and clinical applications, including disease modeling, regenerative medicine, and drug discovery/drug cytotoxicity studies.
组织工程致力于在体外创建具有不同细胞类型的器官功能组件。其中一个挑战是在生理条件下制造用于三维（3D）细胞培养的支架。特别感兴趣的是使用这种支架培养的中枢神经系统的形态和功能的研究。在这里，我们使用弹性体-聚二甲基硅氧烷（PDMS）通过光刻定义的模板来制造格子型支架。具有反点阵列的光掩模通过厚的光刻胶层旋涂，并以 45°的角度向下安装在旋转台上。在重复曝光三次或更多次后，保持相同的曝光计划但旋转相同的角度，开发光刻胶以产生 3D 多孔模板。之后，将 PDMS 的预聚物混合物倒入并固化，然后进行光刻胶刻蚀，得到格子型 PDMS 结构。在细胞培养之前，对 PDMS 格子进行表面功能化。使用 NIH-3T3 细胞和大鼠海马原代细胞进行培养测试，显示出均匀的细胞渗透和 3D 附着。不出所料，与 2D 培养相比，在 3D PDMS 格子中发现了更多的细胞。我们还发现，由于支架形貌和 PDMS 微格子的弹性性质的变化，与 2D 培养相比，3D 培养中的神经元-星形胶质细胞比例更高，细胞分支程度更高。我们的结果表明，3D PDMS 微格子可以提高细胞的存活率和生长，以及神经元的网络形成。我们相信这种使能技术对于研究和临床应用是有用的，包括疾病建模、再生医学和药物发现/药物细胞毒性研究。
