Chang Shu-Han, Ko Ping-Liang, Liao Wei-Hao, Peng Chien-Chung, Tung Yi-Chung
Institute of Biophotonics, National Yang-Ming University, Taipei 11221, Taiwan.
Institute of Biophotonics, National Yang Ming Chiao Tung University, Taipei 11221, Taiwan.
Micromachines (Basel). 2021 Apr 6;12(4):406. doi: 10.3390/mi12040406.
This paper reports a transwell insert-embedded microfluidic device capable of culturing cells at an air-liquid interface (ALI), mimicking the in vivo alveolar epithelium microenvironment. Integration of a commercially available transwell insert makes the device fabrication straightforward and eliminates the tedious device assembly processes. The transwell insert can later be detached from the device for high-resolution imaging of the cells. In the experiments, the cells showing type-I pneumocyte markers are exploited to construct an in vitro alveolar epithelium model, and four culture conditions including conventional liquid/liquid culture (LLC) and air-liquid interface (ALI) cell culture in normal growth medium, and ALI cell culture with inflammatory cytokine (TNF-α) stimulation and ethanol vapor exposure are applied to investigate their effects on the alveolar epithelium barrier function. The barrier permeability is time-lapse monitored using trans-epithelial electrical resistance (TEER) measurement and immunofluorescence staining of the tight junction protein (ZO-1). The results demonstrate the functionalities of the device, and further show the applications and advantages of the constructed in vitro cell models for the lung studies.
本文报道了一种插入Transwell小室的微流控装置，该装置能够在气液界面（ALI）培养细胞，模拟体内肺泡上皮微环境。集成市售的Transwell小室使装置制造变得简单，并省去了繁琐的装置组装过程。之后可将Transwell小室从装置上拆卸下来，用于细胞的高分辨率成像。在实验中利用显示I型肺上皮细胞标志物的细胞构建体外肺泡上皮模型，并应用四种培养条件，包括在正常生长培养基中的传统液/液培养（LLC）和气液界面（ALI）细胞培养，以及炎性细胞因子（TNF-α）刺激和乙醇蒸汽暴露下的ALI细胞培养，以研究它们对肺泡上皮屏障功能的影响。使用跨上皮电阻（TEER）测量和紧密连接蛋白（ZO-1）的免疫荧光染色对屏障通透性进行实时监测。结果证明了该装置的功能，并进一步展示了所构建的体外细胞模型在肺部研究中的应用和优势。
