microfluidic perfusion of huvec and c2bbe1 cultured biochips for analyzing immune responses to microbial colonization

Enhanced Intestine-on-Chip Platform for Immunological Insight

Organ-on-Chip (OoC) systems represent an emerging technique of 3D cell culture that is capable of bridging the gap between conventional 2D cell culture and animal models. OoC platforms typically consist of one or more compartments containing tissue-specific cells grown on a wide range of scaffolds such as membranes or hydrogels1. The models are capable of mimicking one or more defined organotypic functions. Pumps enable continuous microfluidic perfusion of cell culture medium for removal of cellular waste products, supply with nutrition and growth factors for improved cellular differentiation, and recreating essential in vivo conditions. With the integration of immune cells, OoC systems can mimic human immune response in vitro2. To date, a wide range of organs and functional units have been presented1. These systems include models of the vasculature3, lung4, liver2,5, and intestine6 that can be facilitated for drug testing5,7 and infection studies6,8.
We here present a human intestine-on-chip model integrating human epithelial cells forming an organotypic 3D topography of villus-like and crypt-like structures combined with an endothelial lining and tissue-resident macrophages. The model is cultured in a microfluidically perfused biochip in the format of a microscopic slide.&#160;Each biochip consists of two separate microfluidic cavities. Each cavity is divided by a porous polyethylene terephthalate (PET) membrane into an upper and lower chamber. The membrane itself also serves as the scaffold for the cells to grow on each side. The pores of the membrane enable cellular crosstalk and cell migration between cell layers. Each chamber can be accessed by two female luer lock-sized ports. Optionally, an additional mini-luer lock-sized port can provide access to the upper or lower chamber (Figure 1).
The OoC platform offers a number of readouts that can be obtained from a single experiment. The intestine-on-chip is tailored towards combining perfused 3D cell culture, effluent analysis, and fluorescence microscopy to assess cell marker expression, metabolization rates, immune response, microbial colonization and infection, and barrier function3,6,8. The model includes tissue-resident immune cells and direct contact of living microorganisms with the host tissue, which is a benefit compared to other published models9. Further, epithelial cells self-organize into three-dimensional structures that provide a physiologically relevant interface for the colonization with a living microbiota6.

Author Spotlight: Developing Immunocompetent Organ-on-Chip Models for Infectious Disease Research

Immunocompetent Intestine-on-Chip Model for Analyzing Gut Mucosal Immune Responses

Our research focuses on the development and use of immunocompetent organ-on-chip platforms. By creating these physiologically relevant, humanized in vitro models, we are able to study the complex host-pathogen interaction. The knowledge we create can be used to manipulate and identify molecular targets for the therapy of infectious diseases.One key challenge is maintaining a living microbiota under homeostatic conditions alongside host tissue in vitro. This includes replicating the organotypic 3D topography of crypt and villus structures and their microbial colonization. Intestine-on-chip platforms address this by using flow to shape the organotypic structures and prevent bacterial overgrowth.This ensures stable culture conditions. Compared to other available intestine-on-chip platforms, our model enables the detailed study of the human immune response to pathogens due to the inclusion of primary tissue-resident immune cells. Additionally, the model size enables multiple readouts in one single experiment, reducing the overall cost and number of experiments.Our laboratory is focusing on using induced pluripotent stem cells to develop autologous systems that enable personalized medicine.

Microfluidic Perfusion of HUVEC and C2BBe1 Cultured Biochips for Analyzing Immune Responses to Microbial Colonization

microfluidic-perfusion-huvec-c2bbe1-cultured-biochips-for-analyzing

Research

JoVE Journal

Immunology and Infection

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