Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, Nebraska, USA.
Biomater Sci. 2018 Dec 18;7(1):373-388. doi: 10.1039/c8bm01095a.
Human pluripotent stem cell derived endothelial cells (hPSC-ECs) are of great value for studying and treating vascular diseases. However, manufacturing high quantity and quality hPSC-ECs with current cell culture technologies remains very challenging. Here, we report a novel method that can manufacture hPSC-ECs in scalable and cell-friendly microenvironments to address this challenge. Using this method, hPSCs are expanded and differentiated into ECs in microscale alginate hydrogel tubes. The hydrogel tubes protect cells from the highly variable hydrodynamic conditions and critical hydrodynamic stresses in the culture vessel and limit the cell mass less than the diffusion limits (of human tissue) to ensure efficient mass transport. The hydrogel tubes provide uniform and friendly microenvironments for cells to grow. This novel design leads to extremely high production efficiency. We showed that hPSC-ECs could be produced in 10 days with high viability (>90%), high purity (>80%) and high yield (∼5.0 × 108 cells per mL of microspace). The yield is about 250 times that of the current-state-of-the-art. hPSC-ECs made in these hydrogel tubes had similar in vitro and in vivo functions to hPSC-ECs generated by conventional cell culture methods. This simple, scalable, efficient, defined and cost-effective technology will make hPSC-ECs broadly available and affordable for various biomedical applications.
人多能干细胞衍生的内皮细胞（hPSC-ECs）对于研究和治疗血管疾病具有重要价值。然而，用当前的细胞培养技术制造高质量和高数量的 hPSC-ECs 仍然极具挑战性。在这里，我们报告了一种新方法，可以在可扩展和细胞友好的微环境中制造 hPSC-ECs，以解决这一挑战。使用这种方法，hPSCs 在微尺度海藻酸盐水凝胶管中被扩增并分化为 ECs。水凝胶管保护细胞免受培养容器中高度变化的流体动力学条件和关键流体动力学应力的影响，并将细胞质量限制在小于扩散极限（人体组织）的范围内，以确保有效的质量传递。水凝胶管为细胞提供均匀和友好的微环境进行生长。这种新颖的设计导致极高的生产效率。我们表明，hPSC-ECs 可以在 10 天内以高活力（>90%）、高纯度（>80%）和高产率（∼5.0×108 个细胞/mL 微空间）生产。产量约为当前最先进水平的 250 倍。这些水凝胶管中制造的 hPSC-ECs 在体外和体内功能上与通过常规细胞培养方法生成的 hPSC-ECs 相似。这项简单、可扩展、高效、定义明确且具有成本效益的技术将使 hPSC-ECs 广泛可用且负担得起，适用于各种生物医学应用。
