Design of biomimetic substrates for long-term maintenance of alveolar epithelial cells
James C. H. Poon, Zhongfa Liao, Takaya Suzuki, Miranda M. Carleton, John P. Soleas, J. Stewart Aitchison, Golnaz Karoubi, Alison P. McGuigan, Thomas K. Waddell
文献索引：10.1039/C7BM00647K
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摘要
There is a need to establish in vitro lung alveolar epithelial culture models to better understand the fundamental biological mechanisms that drive lung diseases. While primary alveolar epithelial cells (AEC) are a useful option to study mature lung biology, they have limited utility in vitro. Cells that survive demonstrate limited proliferative capacity and loss of phenotype over the first 3–5 days in traditional culture conditions. To address this limitation, we generated a novel physiologically relevant cell culture system for enhanced viability and maintenance of phenotype. Here we describe a method utilizing e-beam lithography, reactive ion etching, and replica molding to generate poly-dimethylsiloxane (PDMS) substrates containing hemispherical cavities that mimic the architecture and size of mouse and human alveoli. Primary AECs grown on these cavity-containing substrates form a monolayer that conforms to the substrate enabling precise control over cell sheet architecture. AECs grown in cavity culture conditions remain viable and maintain their phenotype over one week. Specifically, cells grown on substrates consisting of 50 μm diameter cavities remained 96 ± 4% viable and maintained expression of surfactant protein C (SPC), a marker of type 2 AEC over 7 days. While this report focuses on primary lung alveolar epithelial cells, our culture platform is potentially relevant and useful for growing primary cells from other tissues with similar cavity-like architecture and could be further adapted to other biomimetic shapes or contours.