Ultrathin transparent membranes for cellular barrier and co-culture models.
Ultrathin transparent membranes for cellular barrier and co-culture models.
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DOI：
10.1088/1758-5090/aa5ba7
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发表时间：
2017-02-14
期刊：
影响因子：
9
通讯作者：
Gaborski TR
中科院分区：
文献类型：
--
作者：
Carter RN;Casillo SM;Mazzocchi AR;DesOrmeaux JS;Roussie JA;Gaborski TR
典型的体外屏障和共培养模型依赖于物理上分隔两个隔室的厚的半渗透聚合物膜。聚合物径迹蚀刻膜虽然可渗透小分子，但在与共培养细胞的物理相互作用方面远离生理学，并且由于光散射和自发荧光而与高分辨率成像不相容。在这里，我们报告的光学透明的多孔膜的孔隙率超过20%。我们优化了沉积和退火条件，以产生拉伸和坚固的多孔二氧化硅膜，其厚度与血管基底膜（100-300 nm）相当。我们证明，人脐静脉内皮细胞（HUVECs）的传播和增殖这些膜上类似于控制基板。此外，当HUVEC在膜的相对侧上共培养时，HUVEC能够将细胞质货物转移到脂肪来源的干细胞，证明其厚度支持生理相关的细胞相互作用。最后，我们确认，这些多孔玻璃膜与剥离工艺兼容，产生具有许多平方厘米的活性面积的膜片。我们相信，这些膜将使新的体外屏障和共培养模型，同时提供显着改善的可视化相比，传统的替代品。
Typical in vitro barrier and co-culture models rely upon thick semi-permeable polymeric membranes that physically separate two compartments. Polymeric track-etched membranes, while permeable to small molecules, are far from physiological with respect to physical interactions with co-cultured cells and are not compatible with high-resolution imaging due to light scattering and autofluorescence. Here we report on an optically transparent ultrathin membrane with porosity exceeding 20%. We optimize deposition and annealing conditions to create a tensile and robust porous silicon dioxide membrane that is comparable in thickness to the vascular basement membrane (100–300 nm). We demonstrate that human umbilical vein endothelial cells (HUVECs) spread and proliferate on these membranes similarly to control substrates. Additionally, HUVECs are able to transfer cytoplasmic cargo to adipose-derived stem cells when they are co-cultured on opposite sides of the membrane, demonstrating its thickness supports physiologically relevant cellular interactions. Lastly, we confirm that these porous glass membranes are compatible with lift-off processes yielding membrane sheets with an active area of many square centimeters. We believe that these membranes will enable new in vitro barrier and co-culture models while offering dramatically improved visualization compared to conventional alternatives.