Department of Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea.
Department of Physiology, Yonsei University College of Medicine, Seoul 120-752, Republic of Korea.
Biomaterials. 2015 May;50:127-39. doi: 10.1016/j.biomaterials.2015.01.015. Epub 2015 Feb 17.
Current protocols for human pluripotent stem cell (hPSC) expansion require feeder cells or matrices from animal sources that have been the major obstacle to obtain clinical grade hPSCs due to safety issues, difficulty in quality control, and high expense. Thus, feeder-free, chemically defined synthetic platforms have been developed, but are mostly confined to typical polystyrene culture plates. Here, we report a chemically defined, material-independent, bio-inspired surface coating allowing for feeder-free expansion and maintenance of self-renewal and pluripotency of hPSCs on various polymer substrates and devices. Polydopamine (pDA)-mediated immobilization of vitronectin (VN) peptides results in surface functionalization of VN-dimer/pDA conjugates. The engineered surfaces facilitate adhesion, proliferation, and colony formation of hPSCs via enhanced focal adhesion, cell-cell interaction, and biophysical signals, providing a chemically defined, xeno-free culture system for clonal expansion and long-term maintenance of hPSCs. This surface engineering enables the application of clinically-relevant hPSCs to a variety of biomedical systems such as tissue-engineering scaffolds and medical devices.
目前，人类多能干细胞（hPSC）的扩增方案需要来自动物源的饲养细胞或基质，由于安全问题、质量控制困难和费用高昂，这一直是获得临床级 hPSC 的主要障碍。因此，已经开发出无饲养细胞、化学定义的合成平台，但这些平台大多局限于典型的聚苯乙烯培养板。在这里，我们报告了一种化学定义的、与材料无关的、受生物启发的表面涂层，允许在各种聚合物基底和器件上无饲养细胞扩增和维持 hPSC 的自我更新和多能性。通过多巴胺（pDA）介导的纤连蛋白（VN）肽的固定化，实现了 VN-二聚体/pDA 缀合物的表面功能化。通过增强的粘着斑、细胞-细胞相互作用和生物物理信号，工程表面促进 hPSC 的黏附、增殖和集落形成，为 hPSC 的克隆扩增和长期维持提供了一个无动物来源的化学定义培养系统。这种表面工程使临床相关的 hPSC 能够应用于各种生物医学系统，如组织工程支架和医疗器械。
