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首页> 外文期刊>Biomaterials Science >Development of bioactive hydrogel capsules for the 3D expansion of pluripotent stem cells in bioreactors
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Development of bioactive hydrogel capsules for the 3D expansion of pluripotent stem cells in bioreactors

机译：开发用于生物反应器中多能干细胞3D扩展的生物活性水凝胶胶囊

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Pluripotent stem cells hold great promise for many pharmaceutical and therapeutic applications. However, the lack of scalable methodologies to expand these cells to clinically relevant numbers is a major roadblock in realizing their full potential. To address this problem, we report here a scalable approach for the expansion of pluripotent stem cells within bioactive hydrogel capsules in stirred bioreactors. To achieve rapid crosslinking of cellular microenvironments with tuneable, cell-instructive functionality, we combined calcium-mediated alginate (CaAlg) complexation with crosslinking of poly(ethylene glycol) (PEG) macromers via a Michael-type addition. The resulting hybrid networks have been shown to have very good handling properties and can be readily decorated with biologically active signals such as integrin ligands or Cadherin-based motifs to influence the fate of mouse induced pluripotent stem (iPS) cells. Air-driven co-axial extrusion was used to reproducibly generate gel microcapsules in high-throughput. Furthermore, the gel capsules can be enveloped in a poly(L-lysine) shell to control swelling or molecular permeability independently of the gel composition. iPS cells entrapped within such capsules expanded with limited commitment to the endodermal lineage. Functionalization of gels with an appropriate density of Arg-Gly-Asp (RGD) ligands further increased the iPS cell expansion rate and reduced the spontaneous differentiation. Therefore, the combination of micro-scale instruction of cell fate by an engineered microenvironment and macro-scale cell manipulation in bioreactors opens up exciting opportunities for stem cell-based applications.
机译：多能干细胞在许多药物和治疗应用中具有广阔的前景。然而，缺乏可扩展的方法来将这些细胞扩展到临床相关的数目是实现其全部潜力的主要障碍。为了解决这个问题，我们在这里报告了一种可扩展的方法，用于在搅拌的生物反应器中的生物活性水凝胶胶囊内扩展多能干细胞。为了实现具有可调节的细胞指导功能的细胞微环境的快速交联，我们通过迈克尔型加成结合了钙介导的藻酸盐（CaAlg）络合物与聚（乙二醇）（PEG）大分子单体的交联。所得的杂种网络已显示具有非常好的处理性能，并且可以容易地用诸如整联蛋白配体或基于钙黏着蛋白的基序等生物活性信号修饰，从而影响小鼠诱导的多能干（iPS）细胞的命运。空气驱动的同轴挤出用于可重现地生成高通量的凝胶微胶囊。此外，可以将凝胶胶囊包裹在聚（L-赖氨酸）壳中以独立于凝胶组成来控制溶胀或分子渗透性。包埋在此类胶囊中的iPS细胞以对内胚层谱系的有限承诺进行扩增。具有适当密度的Arg-Gly-Asp（RGD）配体的凝胶功能化进一步提高了iPS细胞的扩增速率并降低了自发分化。因此，在生物反应器中通过工程化微环境进行的细胞命运的微观尺度指示与大规模细胞操纵相结合，为基于干细胞的应用提供了令人兴奋的机会。

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