École Normale Supérieure-PSL Research University, Département de Chimie, Sorbonne Universités-UPMC Univ Paris 06, CNRS UMR 8640 PASTEUR, 24, rue Lhomond, F-75005 Paris, France.
Biofabrication. 2018 Oct 30;11(1):015005. doi: 10.1088/1758-5090/aae0a5.
Substrate elasticity regulates cell functions including cell aggregation and stem cell differentiation. The ability to manufacture substrates of desired elasticity over a broad range is therefore crucial for both fundamental research and advanced applications. In this work, we developed a method to fabricate dense elastomer pillars of different heights on a rigid substrate, providing an effective elasticity ranging from 3 to 168 kPa. Assisted with an elastomer stencil of honeycomb pattern for cell seeding, we obtained uniform colonies of human induced pluripotent stem cells (hiPSCs) and differentiated cardiomyocytes on the pillar substrates of different modulus. Our results showed that the elasticity of substrates significantly affected the cell colony formation via governing the colony edge propagation. More importantly, the results demonstrated that an intermediate substrate elasticity of about 9 kPa is preferable to reach an embryoid-like aggregation and optimal for cardiac differentiation of hiPSCs. Overall, this work sheds new insights on the importance of substrate modulus on cell aggregation and stem cell differentiation as well as the manufacturing of culture substrates with desired elasticity.
基质弹性调节细胞功能，包括细胞聚集和干细胞分化。因此，能够在广泛的范围内制造具有所需弹性的基质对于基础研究和高级应用都至关重要。在这项工作中，我们开发了一种在刚性基底上制造不同高度的密集弹性体支柱的方法，提供了从 3 到 168kPa 的有效弹性范围。借助具有蜂窝图案的弹性体模板进行细胞播种，我们在不同模量的支柱基质上获得了均匀的人诱导多能干细胞（hiPSC）集落和分化的心肌细胞。我们的结果表明，通过控制集落边缘的扩展，基底的弹性显著影响细胞集落的形成。更重要的是，结果表明，约 9kPa 的中间基质弹性有利于达到类胚状体聚集，并最有利于 hiPSC 的心脏分化。总的来说，这项工作揭示了基底模量对细胞聚集和干细胞分化的重要性，以及制造具有所需弹性的培养基底的新见解。
