Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India.
Biomed Mater. 2023 Feb 9;18(2). doi: 10.1088/1748-605X/acb7c0.
Cell-based assays are essentialtools for understanding basic cell biology, pathophysiology of diseases and mechanism of drug actions most cancer studies have utilized two-dimensional (2D) cell culture methods, which have their shortcomings including lack of cell- extracellular matrix interactions and three-dimensional (3D) geometry, and inaccurate representation of cell polarity. Hence, 3D matrices are being increasingly used to study the effect of 3D niche on cell behavior. Till date, very few systematic studies have been done to show comparison of cell behavior when seeded on the surface and encapsulated inside the matrix. In this study, we fabricated poly(ethylene glycol) (PEG) and gelatin-based matrices using UV mediated photo-polymerization to establish 2D and 3D cell culture methods using breast cancer MDA-MB-231 cells. We have found that the adhesion and spreading of cells on the gel surface is different from that when embedded in gels. The stiffness of poly (ethylene glycol) diacrylate (PEGDA)-gelatin methacryloyl (GelMA) hydrogels with lower concentration of GelMA is lower than that with higher GelMA; further, those with higher overall concentration of polymers (>5%) retain their mechanical integrity and do not degrade even after 7 d. Physical characterization of these matrices demonstrate their optimal pore size, mechanical stiffness and degradation, which are further tunable for tissue engineering, regenerative medicine, drug delivery and cancer studies. Additionally, these semi-synthetic PEGDA-GelMA matrices are transparent in nature, thereby, allowing easy imaging of cells in 3D. The system developed here can be used for short and long term cell culture and can be potentially explored for cell migration and metastasis studies.
基于细胞的测定法是理解基础细胞生物学、疾病病理生理学和药物作用机制的重要工具。大多数癌症研究都利用了二维（2D）细胞培养方法，但这些方法存在局限性，例如缺乏细胞-细胞外基质相互作用和三维（3D）几何形状，以及细胞极性的不准确表示。因此，3D 基质越来越多地被用于研究 3D 生态位对细胞行为的影响。迄今为止，很少有系统的研究表明在基质表面和内部接种细胞时细胞行为的比较。在这项研究中，我们使用紫外光介导的光聚合制造了基于聚乙二醇（PEG）和明胶的基质，以建立使用乳腺癌 MDA-MB-231 细胞的 2D 和 3D 细胞培养方法。我们发现，细胞在凝胶表面的粘附和铺展与嵌入凝胶中的情况不同。低浓度明胶甲基丙烯酰化（GelMA）的聚（乙二醇）二丙烯酸酯（PEGDA）-明胶（GelMA）水凝胶的杨氏模量低于高浓度 GelMA 的水凝胶；此外，那些具有更高聚合物总浓度（>5%）的水凝胶保留了其机械完整性，即使在 7 天后也不会降解。这些基质的物理特性表征表明它们具有最佳的孔径、机械硬度和降解性，这些特性可进一步用于组织工程、再生医学、药物输送和癌症研究。此外，这些半合成的 PEGDA-GelMA 基质在本质上是透明的，因此允许在 3D 中轻松成像细胞。这里开发的系统可用于短期和长期细胞培养，并可潜在地用于细胞迁移和转移研究。
