Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Cambridge, Massachusetts 02139, USA.
Lab Chip. 2013 Sep 21;13(18):3569-77. doi: 10.1039/c3lc50252j.
The research areas of tissue engineering and drug development have displayed increased interest in organ-on-a-chip studies, in which physiologically or pathologically relevant tissues can be engineered to test pharmaceutical candidates. Microfluidic technologies enable the control of the cellular microenvironment for these applications through the topography, size, and elastic properties of the microscale cell culture environment, while delivering nutrients and chemical cues to the cells through continuous media perfusion. Traditional materials used in the fabrication of microfluidic devices, such as poly(dimethylsiloxane) (PDMS), offer high fidelity and high feature resolution, but do not facilitate cell attachment. To overcome this challenge, we have developed a method for coating microfluidic channels inside a closed PDMS device with a cell-compatible hydrogel layer. We have synthesized photocrosslinkable gelatin and tropoelastin-based hydrogel solutions that were used to coat the surfaces under continuous flow inside 50 μm wide, straight microfluidic channels to generate a hydrogel layer on the channel walls. Our observation of primary cardiomyocytes seeded on these hydrogel layers showed preferred attachment as well as higher spontaneous beating rates on tropoelastin coatings compared to gelatin. In addition, cellular attachment, alignment and beating were stronger on 5% (w/v) than on 10% (w/v) hydrogel-coated channels. Our results demonstrate that cardiomyocytes respond favorably to the elastic, soft tropoelastin culture substrates, indicating that tropoelastin-based hydrogels may be a suitable coating choice for some organ-on-a-chip applications. We anticipate that the proposed hydrogel coating method and tropoelastin as a cell culture substrate may be useful for the generation of elastic tissues, e.g. blood vessels, using microfluidic approaches.
组织工程和药物开发领域对器官芯片研究表现出了越来越大的兴趣,在器官芯片研究中,可以对与生理或病理相关的组织进行工程改造,以测试药物候选物。微流控技术通过微尺度细胞培养环境的形貌、大小和弹性特性来控制这些应用中的细胞微环境,同时通过连续介质灌注向细胞传递营养物质和化学信号。传统的微流控器件制造材料,如聚二甲基硅氧烷(PDMS),提供了高保真度和高特征分辨率,但不利于细胞附着。为了克服这一挑战,我们开发了一种在封闭的 PDMS 器件内的微流道表面涂覆细胞相容水凝胶层的方法。我们合成了可光交联的明胶和原弹性蛋白基水凝胶溶液,用于在连续流条件下涂覆 50μm 宽的直微流道内的表面,在通道壁上生成水凝胶层。我们观察到在这些水凝胶层上接种的原代心肌细胞表现出对原弹性蛋白涂层的优先附着以及更高的自发搏动率,与明胶相比。此外,在 5%(w/v)的水凝胶涂层通道上,细胞附着、排列和搏动比在 10%(w/v)的水凝胶涂层通道上更强。我们的结果表明,心肌细胞对弹性软的原弹性蛋白培养基底反应良好,表明原弹性蛋白基水凝胶可能是某些器官芯片应用的合适涂层选择。我们预计,所提出的水凝胶涂层方法和原弹性蛋白作为细胞培养基底可能有助于使用微流控方法生成弹性组织,例如血管。