The Contributions of Human Mini-Intestines to the Study of Intestinal Physiology and Pathophysiology

Abstract

The lack of accessibility to normal and diseased human intestine and the inability to separate the different functional compartments of the intestine even when tissue could be obtained have held back the understanding of human intestinal physiology. Clevers and his associates identified intestinal stem cells and established conditions to grow “mini-intestines” ex vivo in differentiated and undifferentiated conditions. This pioneering work has made a new model of the human intestine available and has begun making contributions to the understanding of human intestinal transport in normal physiologic conditions and the pathophysiology of intestinal diseases. However, this model is reductionist and lacks many of the complexities of normal intestine. Consequently, it is not yet possible to predict how great the advances using this model will be for understanding human physiology and pathophysiology, nor how the model will be modified to include multiple other intestinal cell types and physical forces necessary to more closely approximate normal intestine. This review describes recent studies using mini-intestines, which have readdressed previously established models of normal intestinal transport physiology and newly examined intestinal pathophysiology. The emphasis is on studies with human enteroids grown either as three-dimensional spheroids or two-dimensional monolayers. In addition, comments are provided on mouse studies in cases when human studies have not yet been described.

The stress polarity signaling (SPS) pathway serves as a marker and a target in the leaky gut barrier: implications in aging and cancer

“…In this model, crypt-derived stem cells isolated from human or mouse colon (see the Materials and Methods section) were used to generate organoids and later differentiated into polarized enteriod-derived monolayers (EDMs). These EDMs have been validated as model systems that closely resemble the physiologic gut lining in which all cell types (enterocytes, goblet, Paneth enteroendocrine, and tuft cells) are proportionately represented (Sato et al, 2009;Miyoshi & Stappenbeck, 2013;Foulke-Abel et al, 2014;Noel et al, 2017;Yu et al, 2017). We used this "gut-in-a-dish" model to rigorously interrogate the impact of various stressors on the integrity of the gut barrier and cellular processes, using readouts illustrated in Fig 1B. Using pS245-GIV as a marker, we confirmed that the SPSpathway is active in the EDMs, colocalized with the TJ protein, occludin, as previously shown in MDCK cells; (Aznar et al, 2016).…”

A Fast and Simple Fluorometric Method to Detect Cell Death in 3D Intestinal Organoids

“…Intestinal organoids are being used to study not only basic intestinal physiology [6], but also pathophysiological processes, for example, TNFα-induced epithelial cell death during inflammatory bowel disease [7,8]. Moreover, intestinal organoids have been used to study host–pathogen interactions, for example, during Zika virus infection [9].…”

Dysregulation of the engulfment pathway in the gut fuels Inflammatory Bowel Disease

“…To investigate the role of ELMO1 in the IBD-afflicted gut epithelium, we utilized the stem-cell derived 3D intestinal organoids or enteroids 30 . Enteroids mimic the in vivo situation with four different cell types' epithelial, goblet, Paneth cells and enterocytes [31][32][33] . Therefore, the use of human crypt-derived enteroids recreates normal intestinal physiology.…”