Izadifar Zohreh, Ingber Donald E
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.
Department of Urology and Department of Surgery, Harvard Medical School and Boston Children's Hospital, Boston, MA, USA.
Bio Protoc. 2025 Apr 5;15(7):e5262. doi: 10.21769/BioProtoc.5262.
Pathological conditions of the cervix ranging from cervical cancer to structural dysfunction associated with preterm labor all have limited treatment options. Thus, there is a need for physiologically relevant preclinical models that recapitulate the structure and function of this human organ. Here, we describe a protocol for engineering and studying a highly functional in vitro model of the human cervix that is composed of a commercially available, dual-channel, microfluidic, organ-on-a-chip (Organ Chip) device lined by primary cervical epithelial (CE) cells interfaced across a porous membrane with cervical stromal cells. The provision of dynamic and customized media flow through both the epithelial and stromal compartments results in cell growth and differentiation, including the accumulation of a thick mucus layer overlying the epithelium. The resulting model closely mimics the structure, epithelial barrier, mucus composition and structure, and biochemical properties of the in vivo human cervix, as well as its responsiveness to female hormones, pH, and microbiome. This Cervix Chip protocol also includes noninvasive techniques for longitudinal monitoring of the live 3D tissue model. The Cervix Chip offers a powerful preclinical platform for replicating in vivo cervical physiology, studying disease mechanisms, and facilitating the development of new therapeutics and diagnostics. Key features • Creates a functional and physiologically responsive 3D tissue model of the human cervix including a living epithelial-stromal interface. • Enables longitudinal and endpoint analysis of the epithelial and stromal environment and their respective secretions independently. • Allows extended clinically relevant studies, such as assessment of tissue barrier function and mucus production as well as co-culture with microbiome and pathogens. • Uses a commercially available dual-channel microfluidic chip and automated culture system (Zoë Culture Module, Emulate Inc., USA).
从宫颈癌到与早产相关的结构功能障碍等宫颈病理状况，其治疗选择都很有限。因此，需要生理相关的临床前模型来重现这个人体器官的结构和功能。在这里，我们描述了一种构建和研究人宫颈高功能体外模型的方案，该模型由市售的双通道微流控芯片器官（Organ Chip）装置组成，内衬原代宫颈上皮（CE）细胞，通过多孔膜与宫颈基质细胞相互作用。通过上皮和基质隔室提供动态且定制的培养基流动，可导致细胞生长和分化，包括上皮上积累厚厚的黏液层。所得模型紧密模拟体内人宫颈的结构、上皮屏障、黏液组成和结构以及生化特性，以及其对女性激素、pH值和微生物群的反应性。这种宫颈芯片方案还包括用于对活的三维组织模型进行纵向监测的非侵入性技术。宫颈芯片为复制体内宫颈生理学、研究疾病机制以及促进新治疗方法和诊断方法的开发提供了一个强大的临床前平台。关键特性 • 创建人宫颈的功能性和生理反应性三维组织模型，包括活的上皮-基质界面。 • 能够独立地对上皮和基质环境及其各自的分泌物进行纵向和终点分析。 • 允许进行扩展的临床相关研究，如评估组织屏障功能和黏液产生以及与微生物群和病原体的共培养。 • 使用市售的双通道微流控芯片和自动化培养系统（Zoë培养模块，美国Emulate公司）。
