Development of a Human Neuromuscular Tissue-on-a-Chip Model on a 24-Well-Plate-Format Compartmentalized Microfluidic Device
Development of a Human Neuromuscular Tissue-on-a-Chip Model on a 24-Well-Plate-Format Compartmentalized Microfluidic Device
复制标题
在 24 孔板格式隔室化微流体装置上开发人类神经肌肉组织芯片模型
DOI：
10.1039/d1lc00048a
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发表时间：
2021
期刊：
影响因子：
6.1
通讯作者：
Shimizu K
中科院分区：
文献类型：
--
作者：
Yamamoto K;Yamaoka N;Imaizumi Y;Nagashima T;Furutani T;Ito T;Okada Y;Honda H;Shimizu K
神经肌肉组织的工程化三维模型有望在体外模拟它们的无序状态。虽然已经建立了几个模型，但在体内模拟运动单位中运动神经元(MNS)和骨骼肌(SKM)纤维的物理分离结构仍然具有挑战性。在这项研究中，我们的目标是开发用于MNS和工程化SKM组织精确分区共培养的微型设备。所研制的微型装置可以容纳24个孔板，有一个用于MNS的腔室和一个用于SKM组织的腔室。这两个腔室由轴突的微隧道连接，允许轴突，但不允许连接细胞体。人IPSC(HiPSC)来源的MN球体在一个小室中延长其轴突进入微隧道，到达SKM小室中的组织工程化人SKM，并与肌肉纤维形成功能性神经肌肉连接。在该装置上与MNS共培养的SKM组织对MNS的自发放电做出自发收缩的反应。在MN小室中加入神经递质谷氨酸，可引起共同培养的SKM组织收缩。选择性加入河豚毒素或维库溴铵均可引起SKM的组织松弛，其机制可用抑制机制来解释。我们还演示了在设备上对共培养组织的轴突中部施加化学或机械刺激。因此，在该装置上构建的分区神经肌肉组织模型可以用于表型筛选，以评估候选药物的细胞类型特异性疗效，并将成为治疗神经肌肉疾病的基础研究和药物开发的有用工具。
Engineered three-dimensional models of neuromuscular tissues are promising for use in mimicking their disorder states in vitro. Although several models have been developed, it is still challenging to mimic the physically separated structures of motor neurons (MNs) and skeletal muscle (SkM) fibers in the motor units in vivo. In this study, we aimed to develop microdevices for precisely compartmentalized coculturing of MNs and engineered SkM tissues. The developed microdevices, which fit a well of 24 well plates, had a chamber for MNs and chamber for SkM tissues. The two chambers were connected by microtunnels for axons, permissive to axons but not to cell bodies. Human iPSC (hiPSC)-derived MN spheroids in one chamber elongated their axons into microtunnels, which reached the tissue-engineered human SkM in the SkM chamber, and formed functional neuromuscular junctions with the muscle fibers. The cocultured SkM tissues with MNs on the device contracted spontaneously in response to spontaneous firing of MNs. The addition of a neurotransmitter, glutamate, into the MN chamber induced contraction of the cocultured SkM tissues. Selective addition of tetrodotoxin or vecuronium bromide into either chamber induced SkM tissue relaxation, which could be explained by the inhibitory mechanisms. We also demonstrated the application of chemical or mechanical stimuli to the middle of the axons of cocultured tissues on the device. Thus, compartmentalized neuromuscular tissue models fabricated on the device could be used for phenotypic screening to evaluate the cellular type specific efficacy of drug candidates and would be a useful tool in fundamental research and drug development for neuromuscular disorders.