Multipotent neural stem cells (NSCs) are widely applied in pre-clinical and clinical trials as a cell source to promote tissue regeneration in neurodegenerative diseases. Frequently delivered as dissociated cells, aggregates or self-organized rosettes, it is unknown whether disruption of the NSC rosette morphology or method of formation affect signaling profiles of these cells that may impact uniformity of outcomes in cell therapies. Here we generate a neural cell–cell interaction microchip (NCCIM) as an in vitro platform to simultaneously track an informed panel of cytokines and co-evaluate cell morphology and biomarker expression coupled to a sandwich ELISA platform. We apply multiplex in situ tagging technology (MIST) to evaluate ten cytokines (PDGF-AA, GDNF, BDNF, IGF-1, FGF-2, IL-6, BMP-4, CNTF, β-NGF, NT-3) on microchips for EB-derived rosettes, single cell dissociated rosettes and reformed rosette neurospheres. Of the cytokines evaluated, EB-derived rosettes secrete PDGF-AA, GDNF and FGF-2 prominently, whereas this profile is temporarily lost upon dissociation to single cells and in reformed neurospheres two additional cytokines, BDNF and β-NGF, are also secreted. This study on NSC rosettes demonstrates the development, versatility and utility of the NCCIM as a sensitive multiplex detector of cytokine signaling in a high throughput and controlled microenvironment. The NCCIM is expected to provide important new information to refine cell source choices in therapies as well as to support development of informative 2D or 3D in vitro models including areas of neurodegeneration or neuroplasticity.

中文翻译：

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通过新型神经细胞-细胞相互作用微芯片对神经细胞因子信号进行多重分析。

多能神经干细胞（NSCs）被广泛应用于临床前和临床试验，作为促进神经退行性疾病组织再生的细胞来源。经常以解离细胞、聚集体或自组织玫瑰花结的形式提供，目前尚不清楚 NSC 玫瑰花结形态的破坏或形成方法是否会影响这些细胞的信号传导谱，从而影响细胞治疗结果的一致性。在这里，我们生成了一个神经细胞 - 细胞相互作用微芯片 (NCCIM) 作为体外平台，以同时跟踪一组知情的细胞因子并共同评估与夹心 ELISA 平台耦合的细胞形态和生物标志物表达。我们在原位应用多重标记技术 (MIST) 在微芯片上评估 EB 衍生的 10 种细胞因子（PDGF-AA、GDNF、BDNF、IGF-1、FGF-2、IL-6、BMP-4、CNTF、β-NGF、NT-3）玫瑰花结、单细胞解离玫瑰花结和重新形成的玫瑰花结神经球。在评估的细胞因子中，EB 衍生的玫瑰花结显着地分泌 PDGF-AA、GDNF 和 FGF-2，而这种特征在分离成单个细胞后暂时丢失，在重组的神经球中，还分泌了两种额外的细胞因子，BDNF 和 β-NGF。这项关于 NSC 玫瑰花结的研究证明了 NCCIM 作为高通量和受控微环境中细胞因子信号的灵敏多重检测器的开发、多功能性和实用性。NCCIM 有望提供重要的新信息，以完善治疗中的细胞来源选择，并支持信息丰富的 2D 或 3D 的开发体外模型，包括神经变性或神经可塑性领域。