ACS Biomater Sci Eng. 2024 May 13;10(5):3280-3292. doi: 10.1021/acsbiomaterials.4c00312. Epub 2024 Apr 12.
Hydrogels have gained significant popularity as model platforms to study reciprocal interactions between cells and their microenvironment. While hydrogel tools to probe many characteristics of the extracellular space have been developed, fabrication approaches remain challenging and time-consuming, limiting multiplexing or widespread adoption. Thus, we have developed a modular fabrication approach to generate distinct hydrogel microenvironments within the same 96-well plate for increased throughput of fabrication as well as integration with existing high-throughput assay technologies. This approach enables hydrogel mechanical characterization and is used to generate both elastic and viscoelastic hydrogels across a range of stiffnesses. Additionally, this fabrication method enabled a 3-fold reduction in polymer and up to an 8-fold reduction in fabrication time required per hydrogel replicate. The feasibility of this platform for two-dimensional (2D) cell culture applications was demonstrated by measuring both population-level and single-cell-level metrics microplate reader and high-content imaging. Finally, a 96-well hydrogel array was utilized for three-dimensional (3D) cell culture, demonstrating the ability to support high cell viability. Together, this work demonstrates a versatile and easily adaptable fabrication approach that can support the ever-expanding tool kit of hydrogel technologies for cell culture applications.
水凝胶作为研究细胞与其微环境之间相互作用的模型平台已经得到了广泛的关注。虽然已经开发出了许多用于探测细胞外空间特性的水凝胶工具，但制造方法仍然具有挑战性且耗时，限制了其多重检测或广泛应用。因此，我们开发了一种模块化的制造方法，可以在同一个 96 孔板中生成不同的水凝胶微环境，从而提高制造的通量并与现有的高通量分析技术集成。这种方法能够进行水凝胶力学特性的表征，并用于生成一系列弹性和粘弹性水凝胶，其弹性模量范围很广。此外，这种制造方法使聚合物的用量减少了 3 倍，每个水凝胶复制品的制造时间减少了 8 倍。通过使用微孔板读数器和高内涵成像来测量群体水平和单细胞水平的指标，证明了该平台在二维（2D）细胞培养应用中的可行性。最后，使用 96 孔水凝胶阵列进行三维（3D）细胞培养，证明了其支持高细胞活力的能力。总之，这项工作展示了一种通用且易于适应的制造方法，可以支持不断扩展的水凝胶技术工具包，用于细胞培养应用。
