Structurally and mechanically tuned macroporous hydrogels for scalable mesenchymal stem cell-extracellular matrix spheroid production

• 成果类型: Article
• 署名作者: Yin, Sheng; Wu, Haipeng; Huang, Yaying; Lu, Chenjing; Cui, Jian; Li, Ying; Xue, Bin; Wu, Junhua; Jiang, Chunping; Gu, Xiaosong; Wang, Wei; Cao, Yi
• 署名单位: Nanjing University; Collaborative Innovation Center of Advanced Microstructures (CICAM); Jinan Microecological Biomedicine Shandong Laboratory; Nanjing University of Information Science & Technology; Nanjing University; Nanjing University; Nanjing University; Nanjing University; Nanjing University
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-10669
• DOI: 10.1073/pnas.2404210121
• 发表日期: 2024-07-09
• 关键词: human bone-marrow; osteogenic differentiation; in-vitro; stromal cells; ecm; tgf-beta-3; scaffolds; quiescence; migration; delivery

摘要

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.