Abstract：
Mesenchymal stem cells (MSCs) are essential in regenerative medicine. However, conventional expansion and harvesting methods often fail to maintain the essential extracellular matrix (ECM) components, which are crucial for their functionality and efficacy in therapeutic applications. Here, we introduce a bone marrow-inspired macroporous hydrogel designed for the large-scale production of MSC-ECM spheroids. Through a soft-templating approach leveraging liquid-liquid phase separation, we engineer macroporous hydrogels with customizable features, including pore size, stiffness, bioactive ligand distribution, and enzyme-responsive degradability. These tailored environments are conducive to optimal MSC proliferation and ease of harvesting. We find that soft hydrogels enhance mechanotransduction in MSCs, establishing a standard for hydrogel-based 3D cell culture. Within these hydrogels, MSCs exist as both cohesive spheroids, preserving their innate vitality, and as migrating entities that actively secrete functional ECM proteins. Additionally, we also introduce a gentle, enzymatic harvesting method that breaks down the hydrogels, allowing MSCs and secreted ECM to naturally form MSC-ECM spheroids. These spheroids display heightened stemness and differentiation capacity, mirroring the benefits of a native ECM milieu. Our research underscores the significance of sophisticated materials design in nurturing distinct MSC subpopulations, facilitating the generation of MSC-ECM spheroids with enhanced therapeutic potential.
摘要：
间充质干细胞（MSCs）在再生医学中是必不可少的。然而,传统的扩增和收获方法往往不能保持必要的细胞外基质（ECM）成分，这对它们在治疗应用中的功能和功效至关重要。这里,我们介绍了一种设计用于大规模生产MSC-ECM球体的受骨髓启发的大孔水凝胶。通过利用液-液相分离的软模板方法，我们设计了具有可定制功能的大孔水凝胶，包括孔径，刚度,生物活性配体分布，和酶响应降解性。这些定制的环境有利于最佳MSC增殖和易于收获。我们发现软水凝胶增强MSCs的机械转导，建立基于水凝胶的3D细胞培养标准。在这些水凝胶中，MSCs以两种粘性球体的形式存在，保持他们天生的活力，以及作为积极分泌功能性ECM蛋白的迁移实体。此外,我们还介绍了一个温柔的，分解水凝胶的酶促收获方法，允许MSC和分泌的ECM自然形成MSC-ECM球体。这些球体显示出增强的干性和分化能力，反映原生ECM环境的好处。我们的研究强调了复杂材料设计在培育不同MSC亚群中的重要性，促进具有增强治疗潜力的MSC-ECM球体的生成。
