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Stable chemical bonding of porous membranes and poly(dimethylsiloxane) devices forlong-term cell culture

机译:多孔膜和聚二甲基硅氧烷装置的稳定化学键合长期细胞培养

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摘要

We have investigated the bonding stability of various silane treatments for the integration of track-etched membranes with poly(dimethylsiloxane) (PDMS) microfluidic devices. We compare various treatments using trialkoxysilanes or dipodal silanes to determine the effect of the organofunctional group, cross-link density, reaction solvent, and catalyst on the bond stability. We find that devices made using existing silane methods delaminated after one day when immersed in cell culture medium at 37 °C. In contrast, the dipodal silane, bis[3-(trimethoxysilyl)propyl]amine, is shown to yield stable and functional integration of membranes with PDMS that is suitable for long-term cell culture. To demonstrate application of the technique, we fabricated an open-surface device in which cells cultured on a track-etched membrane can be stimulated at their basal side via embedded microfluidic channels. C2C12 mouse myoblasts were differentiated into myotubes over the course of two weeks on these devices to demonstrate biocompatibility. Finally, devices were imaged during the basal-side delivery of a fluorescent stain to validate the membrane operation and long-term stability of the bonding technique.
机译:我们已经研究了各种硅烷处理的粘结稳定性,以便将径迹蚀刻膜与聚二甲基硅氧烷(PDMS)微流体装置集成在一起。我们比较了使用三烷氧基硅烷或二脚烷硅烷的各种处理方法,以确定有机官能团,交联密度,反应溶剂和催化剂对键稳定性的影响。我们发现,将现有的硅烷方法制成的设备在浸入37°C的细胞培养基中一天后会分层。相反,二足硅烷,双[3-(三甲氧基甲硅烷基)丙基]胺,显示出膜和PDMS膜的稳定和功能整合,适用于长期细胞培养。为了证明该技术的应用,我们制造了一种开放表面的设备,其中可以通过嵌入的微流体通道在其底侧刺激在轨迹蚀刻膜上培养的细胞。在这些设备上,经过两周的过程,将C2C12小鼠成肌细胞分化为肌管,以证明其生物相容性。最后,在荧光染料的基侧传递过程中对设备成像,以验证膜操作和键合技术的长期稳定性。

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