Department of Materials Science and Engineering, University of Washington, Seattle, WA, 98195, USA.
Department of Neurological Surgery, University of Washington, Seattle, WA, 98195, USA.
Macromol Biosci. 2023 Jun;23(6):e2200460. doi: 10.1002/mabi.202200460. Epub 2023 Mar 25.
Human-induced pluripotent stem cells (hiPSCs) cultured in 3D matrices hold great promise in disease modeling, drug discovery, and tissue regeneration. Uniform cell distribution in a 3D structure is critical to the growth and function of hiPSCs, yet cell seeding in 3D matrices often remains superficial, leading to limited cell proliferation and compromised pluripotency. Here, an approach to improve cell penetration depth of hiPSCs in 3D scaffolds modified with hiPSCs conditioned medium (CM) is reported. It is shown that extracellular matrix components are successfully deposited onto the scaffold wall surface after CM treatment and promoted homogeneous cell adhesion during initial seeding. Compared to plain, unmodified scaffolds, the CM treated scaffold improves spatial cell distribution uniformity and upregulates pluripotency markers. Notably, the expression of 29 genes associated with 11 signaling pathways participated in the pluripotency maintenance of hiPSCs exhibits >2-fold change in hiPSCs grown in the CM treated scaffolds than 2D counterparts, demonstrating that CM treated scaffolds can support a more primitive and undifferentiated phenotype of hiPSCs. This study introduces a simple and effective method to enhance cell penetration and maintain cell pluripotency in 3D matrices.
人诱导多能干细胞（hiPSCs）在 3D 基质中培养，在疾病建模、药物发现和组织再生方面具有巨大的应用潜力。在 3D 结构中，均匀的细胞分布对于 hiPSCs 的生长和功能至关重要，然而，细胞在 3D 基质中的接种往往仍然很浅，导致细胞增殖有限，多能性受损。在这里，我们报告了一种通过 hiPSCs 条件培养基（CM）修饰 3D 支架来提高 hiPSCs 细胞穿透深度的方法。结果表明，CM 处理后，细胞外基质成分成功沉积在支架壁表面，并在初始接种时促进了均匀的细胞黏附。与普通、未经修饰的支架相比，CM 处理的支架提高了空间细胞分布的均匀性，并上调了多能性标志物。值得注意的是，参与 hiPSCs 多能性维持的 11 条信号通路中的 29 个基因的表达，在 CM 处理的支架中培养的 hiPSCs 中的变化是 2D 对照组的 2 倍以上，表明 CM 处理的支架可以支持 hiPSCs 更原始和未分化的表型。本研究介绍了一种简单有效的方法，可增强 3D 基质中的细胞穿透性并维持细胞多能性。
