Imaging-guided bioreactor for de-epithelialization and long-term cultivation of ex vivo rat trachea.

Recent synergistic advances in organ-on-chip and tissue engineering technologies offer opportunities to create in vitro-grown tissue or organ constructs that can faithfully recapitulate their in vivo counterparts. Such in vitro tissue or organ constructs can be utilized in multiple applications, including rapid drug screening, high-fidelity disease modeling, and precision medicine. Here, we report an imaging-guided bioreactor that allows in situ monitoring of the lumen of ex vivo airway tissues during controlled in vitro tissue manipulation and cultivation of isolated rat trachea. Using this platform, we demonstrated partial removal of the rat tracheal epithelium (i.e., de-epithelialization) without disrupting the underlying subepithelial cells and extracellular matrix. Through different tissue evaluation assays, such as immunofluorescent staining, DNA/protein quantification, and electron beam microscopy, we showed that the epithelium of the tracheal lumen can be effectively removed with negligible disruption in the underlying tissue layers, such as cartilage and blood vessel. Notably, using a custom-built micro-optical imaging device integrated with the bioreactor, the trachea lumen was visualized at the cellular level, and removal of the endogenous epithelium and distribution of locally delivered exogenous cells were demonstrated in situ. Moreover, the de-epithelialized trachea supported on the bioreactor allowed attachment and growth of exogenous cells seeded topically on its denuded tissue surface. Collectively, the results suggest that our imaging-enabled rat trachea bioreactor and localized cell replacement method can facilitate creation of bioengineered in vitro airway tissue that can be used in different biomedical applications. An imaging-enabled rat trachea bioreactor is developed that can permit controllable in vitro epithelium replacement, in situ cell monitoring, and long-term tissue scaffold cultivation

芯片器官和组织工程技术的协同进步为创建体外培养的组织或器官构建体提供了机会，这些构建体能够如实重现其体内对应物。这种体外组织或器官构建体可用于多种应用，包括快速药物筛选、高保真疾病建模和精准医疗。在此，我们报道了一种成像引导的生物反应器，它能够在对离体大鼠气管进行可控的体外组织操作和培养过程中对其管腔进行原位监测。利用该平台，我们展示了在不破坏下方的上皮下细胞和细胞外基质的情况下部分去除大鼠气管上皮（即去上皮化）。通过不同的组织评估分析，如免疫荧光染色、DNA/蛋白质定量和电子束显微镜检查，我们表明气管腔的上皮可以被有效去除，而对下方的组织层（如软骨和血管）的破坏可以忽略不计。值得注意的是，使用与生物反应器集成的定制微型光学成像设备，气管腔在细胞水平上得以可视化，并且原位展示了内源性上皮的去除以及局部递送的外源性细胞的分布。此外，支撑在生物反应器上的去上皮化气管允许局部接种在其裸露组织表面的外源性细胞附着和生长。总体而言，这些结果表明我们的具有成像功能的大鼠气管生物反应器和局部细胞替换方法能够促进创建可用于不同生物医学应用的生物工程体外气道组织。 开发了一种具有成像功能的大鼠气管生物反应器，它能够允许可控的体外上皮替换、原位细胞监测以及长期的组织支架培养。