Department of Chemical and Biomolecular Engineering, University of Nebraska, Lincoln, Nebraska, United States of America. Biomedical Engineering Program, University of Nebraska, Lincoln, Nebraska, United States of America.
Biofabrication. 2018 Feb 1;10(2):025006. doi: 10.1088/1758-5090/aaa6b5.
Human pluripotent stem cells (hPSCs) are required in large numbers for various biomedical applications. However, the scalable and cost-effective culturing of high quality hPSCs and their derivatives remains very challenging. Here, we report a novel and physiologically relevant 3D culture system (called the AlgTube cell culture system) for hPSC expansion and differentiation. With this system, cells are processed into and cultured in microscale alginate hydrogel tubes that are suspended in the cell culture medium in a culture vessel. The hydrogel tubes protect cells from hydrodynamic stresses in the culture vessel and limit the cell mass smaller than 400 μm in diameter to ensure efficient mass transport, creating cell-friendly microenvironments for growing cells. This system is simple, scalable, highly efficient, defined and compatible with the current good manufacturing practices. Under optimized culture conditions, the AlgTubes enabled long-term culture of hPSCs (>10 passages, >50 days) with high cell viability, high growth rate (1000-fold expansion over 10 days per passage), high purity (>95% Oct4+) and high yield (5.0 × 10 cells ml), all of which offer considerable advantages compared to current approaches. Moreover, the AlgTubes enabled directed differentiation of hPSCs into various tissue cells. This system can be readily scaled to support research from basic biological study to clinical development and the future industry-scale production.
人多能干细胞（hPSCs）在各种生物医学应用中都需要大量使用。然而，高质量 hPSCs 及其衍生物的规模化和具有成本效益的培养仍然极具挑战性。在这里，我们报告了一种新颖且与生理相关的 3D 培养系统（称为 AlgTube 细胞培养系统），用于 hPSC 的扩增和分化。在该系统中，细胞被加工成微尺度海藻酸盐水凝胶管，并在培养容器中的细胞培养基中悬浮。水凝胶管可保护细胞免受培养容器中流体动力应力的影响，并将细胞质量限制在直径小于 400μm 以下，以确保有效的质量传递，为细胞生长创造细胞友好的微环境。该系统简单、可扩展、高效、定义明确且与当前良好生产规范兼容。在优化的培养条件下，AlgTube 能够实现 hPSC 的长期培养（>10 代，>50 天），具有高细胞活力、高生长速度（每代 10 天 1000 倍扩增）、高纯度（>95%Oct4+）和高产量（5.0×10 个细胞/ml），与当前方法相比具有明显优势。此外，AlgTube 能够将 hPSC 定向分化为各种组织细胞。该系统易于扩展，可支持从基础生物学研究到临床开发和未来工业规模生产的研究。
