Three-Dimensional Hyaluronic Acid Hydrogel-Based Models for In Vitro Human iPSC-Derived NPC Culture and Differentiation.
Three-Dimensional Hyaluronic Acid Hydrogel-Based Models for In Vitro Human iPSC-Derived NPC Culture and Differentiation.
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DOI：
10.1039/c7tb00721c
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发表时间：
2017-06-07
期刊：
影响因子：
--
通讯作者：
Jiang P
中科院分区：
文献类型：
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作者：
Wu S;Xu R;Duan B;Jiang P
人诱导多能干细胞衍生的神经祖细胞（hiPSC-NPC）被认为是一种有前途的移植细胞来源，并已用于体外器官制造以重现中枢神经系统（CNS）疾病。hiPSC-NPCs体外三维模型的建立及其分化调控对于理解生物学过程、CNS疾病和再生具有重要意义。在这里，我们实现了3D甲基丙烯酸酯化透明质酸（Me-HA）水凝胶与封装的hiPSC-NPC作为体外培养模型，并进一步研究了水凝胶刚度对hiPSC-NPC的细胞行为的作用。我们首先将单分散的hiPSC-NPC包封在软的和硬的Me-HA水凝胶中，并发现hiPSC-NPC逐渐自组装和聚集形成3D球状体。然后，将hiPSC-NPC以球状体的形式装载到Me-HA水凝胶中，以评估它们响应于水凝胶刚性的自发分化。软Me-HA水凝胶包封的hiPSC-NPC显示出稳健的神经突生长，并显示出高水平的自发神经分化。我们进一步将唐氏综合征（DS）患者特异性hiPSC衍生的NPC（DS-NPC）球状体包封在我们的水凝胶内。DS-NPC在软和硬Me-HA水凝胶中均保持优异的细胞活力。类似地，软水凝胶通过显著上调神经成熟标志物促进DS-NPC的神经分化。该研究表明，软基质促进hiPSC-NPC的神经分化，并且具有hiPSC-NPC或DS-NPC的HA基水凝胶是用于CNS疾病研究的有效3D模型。实施刚度可调的基于3D HA的水凝胶模型以控制hiPSC-NPC或DS-NPC的祖细胞性质和神经元分化。
Human induced pluripotent stem cell-derived neural progenitor cells (hiPSC-NPCs) are considered as a promising cell source for transplantation and have been used for organoid fabrication to recapitulate central nervous system (CNS) diseases in vitro. The establishment of three-dimensional (3D) in vitro model with hiPSC-NPCs and control of their differentiation is significantly critical for understanding biological processes and CNS disease and regeneration. Here we implemented 3D methacrylated hyaluronic acid (Me-HA) hydrogels with encapsulation of hiPSC-NPCs as in vitro culture models and further investigated the role of the hydrogel rigidity on the cell behavior of hiPSC-NPCs. We first encapsulated single dispersive hiPSC-NPCs within both soft and stiff Me-HA hydrogel and found that hiPSC-NPCs gradually self-assembled and aggregated to form 3D spheroids. Then, the hiPSC-NPCs were laden into Me-HA hydrogels in the form of spheroids to evaluate their spontaneous differentiation in response to hydrogel rigidity. The soft Me-HA hydrogel-encapsulated hiPSC-NPCs displayed robust neurite outgrowth and showed high levels of spontaneous neural differentiation. We further encapsulated Down Syndrome (DS) patient-specific hiPSC-derived NPCs (DS-NPCs) spheroids within our hydrogels. DS-NPCs remained excellent cell viability in both soft and stiff Me-HA hydrogels. Similarly, soft hydrogels promoted neural differentiation of DS-NPCs by significantly upregulating neural maturation markers. This study demonstrates that soft matrix promotes neural differentiation of hiPSC-NPCs and HA-based hydrogels with hiPSC-NPCs or DS-NPCs are effective 3D models for CNS disease study. Stiffness tunable 3D HA-based hydrogel models were implemented to control the progenitor properties and neuronal differentiation of hiPSC-NPCs or DS-NPCs.