Department of Chemical and Biological Engineering, University of Wisconsin-Madison, 1415 Engineering Drive, Madison, WI, 53705, USA.
Adv Healthc Mater. 2021 Nov;10(21):e2100995. doi: 10.1002/adhm.202100995. Epub 2021 Aug 29.
Spatiotemporally controlled presentation of morphogens and elaborate modulation of signaling pathways elicit pattern formation during development. Though this process is critical for proper organogenesis, unraveling the mechanisms of developmental biology have been restricted by challenges associated with studying human embryos. Human pluripotent stem cells (hPSCs) have been used to model development in vitro, however difficulties in precise spatiotemporal control of the cellular microenvironment have limited the utility of this model in exploring mechanisms of pattern formation. Here, a simple and versatile method is presented to spatially pattern hPSC differentiation in 2-dimensional culture via localized morphogen adsorption on substrates. Morphogens including bone morphogenetic protein 4 (BMP4), activin A, and WNT3a are patterned to induce localized mesendoderm, endoderm, cardiomyocyte (CM), and epicardial cell (EpiC) differentiation from hPSCs and hPSC-derived progenitors. Patterned CM and EpiC co-differentiation allows investigation of intercellular interactions in a spatially controlled manner and demonstrate improved alignment of CMs in proximity to EpiCs. This approach provides a platform for the controlled and systematic study of early pattern formation. Moreover, this study provides a facile approach to generate 2D patterned hPSC-derived tissue structures for modeling disease and drug interactions.
时空控制的形态发生因子呈现和精细调节的信号通路在发育过程中引发模式形成。虽然这个过程对于正常的器官发生至关重要，但由于研究人类胚胎相关的挑战，发育生物学的机制仍未被完全阐明。人类多能干细胞（hPSC）已被用于体外发育模型研究，然而，精确时空控制细胞微环境的困难限制了该模型在探索模式形成机制方面的应用。本研究提出了一种简单而通用的方法，通过在底物上局部吸附形态发生因子来在 2 维培养中对 hPSC 分化进行空间图案化。形态发生因子包括骨形态发生蛋白 4（BMP4）、激活素 A 和 WNT3a 等被图案化以诱导 hPSC 和 hPSC 衍生祖细胞的局部中胚层、内胚层、心肌细胞（CM）和心外膜细胞（EpiC）分化。CM 和 EpiC 的图案化共分化允许以空间控制的方式研究细胞间相互作用，并证明在 EpiC 附近的 CM 排列更加整齐。该方法为早期模式形成的可控和系统研究提供了一个平台。此外，该研究还提供了一种简便的方法来生成用于疾病和药物相互作用建模的 2D 图案化 hPSC 衍生组织结构。
