Department of Bioengineering, Rice University, 6100 Main St, MS-142, Houston, TX, 77005, USA.
Medical Scientist Training Program, Baylor College of Medicine, One Baylor Plaza, Houston, TX, 77030, USA.
Ann Biomed Eng. 2021 Apr;49(4):1233-1244. doi: 10.1007/s10439-020-02705-8. Epub 2021 Jan 6.
To generate physiologically-relevant experimental models, the study of enteric diarrheal diseases is turning increasingly to advanced in vitro models that combine ex vivo, stem cell-derived "organoid" cell lines with bioengineered culture environments that expose them to mechanical stimuli, such as fluid flow. However, such approaches require considerable technical expertise with both microfabrication and organoid culture, and are, therefore, inaccessible to many researchers. For this reason, we have developed a perfusion system that is simple to fabricate, operate, and maintain. Its dimensions approximate the volume and cell culture area of traditional 96-well plates and allow the incorporation of fastidious primary, stem cell-derived cell lines with only minimal adaptation of their established culture techniques. We show that infections with enteroaggregative E. coli and norovirus, common causes of infectious diarrhea, in the system display important differences from static models, and in some ways better recreate the pathophysiology of in vivo infections. Furthermore, commensal strains of bacteria can be added alongside the pathogens to simulate the effects of a host microbiome on the infectious process. For these reasons, we believe that this perfusion system is a powerful, yet easily accessible tool for studying host-pathogen interactions in the human intestine.
为了生成与生理相关的实验模型，肠易激性腹泻疾病的研究越来越倾向于使用先进的体外模型，这些模型将离体的、源自干细胞的“类器官”细胞系与生物工程培养环境相结合，使它们暴露于机械刺激下，如流体流动。然而，这些方法需要对微加工和类器官培养都有相当的技术专长，因此，许多研究人员无法使用这些方法。出于这个原因，我们开发了一种灌注系统，它易于制造、操作和维护。它的尺寸与传统的 96 孔板的体积和细胞培养面积相近，并允许在不改变其既定培养技术的情况下，将复杂的原代、源自干细胞的细胞系纳入其中。我们表明，在该系统中感染肠聚集性大肠杆菌和诺如病毒（常见的传染性腹泻病原体）与静态模型相比显示出重要差异，并且在某些方面更好地再现了体内感染的病理生理学。此外，可以将共生菌与病原体一起添加到系统中，以模拟宿主微生物组对感染过程的影响。基于这些原因，我们认为该灌注系统是一种强大但易于使用的工具，可用于研究人类肠道中的宿主-病原体相互作用。
