Combining Human Organoids and Organ-on-a-Chip Technology to Model Intestinal Region-Specific Functionality

• DOI: 10.3791/63724
• Publication Date: 2022-05-06T00:00:15Z
• Authors: Gauri Kulkarni; Athanasia Apostolou; Lorna Ewart; Carolina Lucchesi; Magdalena Kasendra
• Keywords: Organoid; Organ-on-a-Chip; Intestinal mucosa

The intestinal mucosa is a complex physical and biochemical barrier that fulfills myriad of important functions. It enables the transport, absorption, metabolism nutrients xenobiotics while facilitating symbiotic relationship with microbiota restricting invasion microorganisms. Functional interaction between various cell types their environment vital to establish maintain tissue homeostasis. Modeling these interactions integrated physiology in vitro formidable goal potential transform way new therapeutic targets drug candidates are discovered developed. Organoids Organ-on-a-Chip technologies have recently been combined generate human-relevant intestine chips suitable for studying functional aspects pathophysiology vitro. derived from biopsies small (duodenum) large seeded into top compartment an organ chip then successfully expand as monolayers preserving distinct cellular, molecular, features each region. Human tissue-specific microvascular endothelial cells incorporated bottom recreate epithelial-endothelial interface. This novel platform facilitates luminal exposure nutrients, drugs, microorganisms, enabling studies permeability, host-microbe interactions. Here, detailed protocol provided establishment representing human duodenum (duodenum chip) colon (colon chip), subsequent culture under continuous flow peristalsis-like deformations. We demonstrate methods assessing CYP3A4 induction using prototypical inducers substrates. Lastly, we provide step-by-step procedure modeling interferon gamma (IFNγ)-mediated disruption (leaky gut syndrome) chip, including evaluating alteration paracellular changes cytokine secretion, transcriptomic profiling within chip.