The blood–brain barrier (BBB) protects the central nervous system from toxins and pathogens in the blood by regulating permeation of molecules through the barrier interface. In vitro BBB models described to date reproduce some aspects of BBB functionality, but also suffer from incomplete phenotypic expression of brain endothelial traits, difficulty in reproducibility and fabrication, or overall cost. To address these limitations, a 3D BBB model based on a hybrid paper/nanofiber scaffold is described. The cell culture platform utilizes lens paper as a framework to accommodate 3D culture of astrocytes. An electrospun nanofiber layer is coated onto one face of the paper to mimic the basement membrane and support growth of an organized 2D layer of endothelial cells (ECs). Human induced pluripotent stem cell-derived ECs and astrocytes are co-cultured to develop a human BBB model. Morphological and spatial organization of model are validated with confocal microscopy. Measurements of transendothelial resistance and permeability demonstrate the BBB model develops a high-quality barrier and responds to hyperosmolar treatments. RNA-sequencing shows introduction of astrocytes both regulates EC tight junction proteins and improves endothelial phenotypes related to vasculogenesis. This model shows promise as a model platform for future in vitro studies of the BBB.

中文翻译：

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用于构建 3D 血脑屏障模型的混合纳米纤维/纸细胞培养平台

血脑屏障（BBB）通过调节屏障界面的分子渗透来保护中枢神经系统免受血液中毒素和病原体的侵害。迄今为止描述的体外 BBB 模型再现了 BBB 功能的某些方面，但也存在脑内皮特征的不完整表型表达、重现性和制造困难或总体成本问题。为了解决这些限制，描述了基于混合纸/纳米纤维支架的 3D BBB 模型。细胞培养平台利用透镜纸作为框架来适应星形胶质细胞的 3D 培养。将电纺纳米纤维层涂覆在纸张的一个面上，以模拟基底膜并支持有组织的二维内皮细胞 (EC) 层的生长。将人类诱导多能干细胞衍生的 EC 和星形胶质细胞共培养以开发人类 BBB 模型。模型的形态和空间组织通过共焦显微镜进行验证。跨内皮阻力和渗透性的测量表明 BBB 模型形成了高质量的屏障并对高渗治疗有反应。RNA测序显示星形胶质细胞的引入既可以调节EC紧密连接蛋白，又可以改善与血管生成相关的内皮表型。该模型有望成为未来 BBB 体外研究的模型平台。