微米阵列结构聚合物薄膜的制备及其对细胞三维培养的影响

Preparation of microarray-structured polymer film and its effect on 3D cell culture

摘要:

二维 (Two-dimensional，2D) 细胞实验模型是目前研究人类疾病的细胞过程和药物筛选的主流方法。然而，生物细胞的生长受到众多因素的影响，传统的2D细胞培养在精确再现三维组织内细胞的功能方面存在一些障碍。与2D细胞培养相比，三维 (Three-dimensional，3D) 细胞培养体系注重细胞间的接触及细胞-基质间的接触，更接近于生物体的生长环境，更适合于药物筛选、细胞培植等研究。文中通过纳米压印技术制备了不同结构微米阵列聚合物薄膜，并将其应用于293T细胞的培养，通过调节薄膜表面结构、表面接触角成功实现了对生物细胞生长形貌的调控。利用扫描电镜等方法，对比了聚合物薄膜不同微结构、不同表面润湿性对细胞生长形貌的影响，重点关注细胞团的形态变化。结果表明，细胞在亲水性10 μm柱形阵列薄膜上呈现三维生长状态，这种薄膜可能更适用于制备生物细胞3D培养基；疏水性3 μm柱形阵列薄膜适用于体积小、表皮硬的组织细胞的3D培养，对于体积较大的细胞效果差。另外，对于疏水性较强的薄膜，细胞倾向于球状生长，而亲水性较强的薄膜则易贴壁生长。研究结果为微结构薄膜在生物细胞3D培养方面的应用作了初步探索。

Abstract:

The two-dimensional (2D) cell culture model is currently used to study cellular processes and drug screening for human diseases. However, the growth of cells is affected by many factors. For conventional 2D cell culture, many of the difficulties are encountered in accurately replicating the cell function in three-dimensional (3D) tissues. Compared with 2D cell culture, much attention is paid to the cell-to-cell and cell-matrix interactions for 3D cell culture systems, which can more closely mimic the growth environment for cultured cells. Therefore, the 3D cell culture system was more suitable for a variety of applications such as drug screening and cell proliferation. In this work, we prepared microarray-structured polymer films with different geometric structures by nanoimprint lithography and used the films as cell culture platforms for the culture of 293T cells. Through the adjustment of the surface morphology and water contact angle of the prepared films, the regulation of the morphological changes of cell growth was successfully realized. Experimental results demonstrated that the hydrophilic films with 10 μm-pillar microstructure are applicable to 3D cell culture, whereas the hydrophobic films with 3 μm-pillar microstructure are only suitable for 3D culture of cells with a smaller size and stiff cuticular layer. In addition, cells tended to the formation of spheroids on the hydrophobic films, while cells usually adhered to the surface and grew on the hydrophilic films. This work represents further technological progress in the development of 3D cell culture, thereby facilitating future studies of physiologically relevant processes.