Developing an advanced gut on chip model enabling the study of epithelial cell/fibroblast interactions

Abstract

We report on a new gut on chip combining the co-culture of primary epithelial and stromal cells in 3D biomimetic scaffold. Proper segregation of dividing and differentiated cells along the crypt-villus axis was achieved in these unique conditions.

New Stable Cell Lines Derived from the Proximal and Distal Intestine of Rainbow Trout (Oncorhynchus mykiss) Retain Several Properties Observed In Vivo

“…Consistent with their epithelial origin, RTgutGC were already known to express zonula occludens 1 [ 18 , 22 , 23 , 26 , 27 ], and we confirmed the expression of this tight junction protein in both RTdi-MI and RTpi-MI. We extended the epithelial characterization of all these lines, showing the expression of E-cadherin and Claudin, two molecules that interact with zonula occludens 1 for maintenance of the intestinal epithelium barrier and signal transduction between adjacent cells [ 28 , 29 ]. RTgutGC cells translate the expression of epithelial barrier genes into the formation of a functional barrier, as indicated by increasing TEER values measured at regular intervals up to 21 days after seeding [ 18 , 23 , 30 , 31 , 32 ].…”

“…Vimentin and collagen type 1 are typically expressed by fibroblasts and myofibroblasts obtained from the human [ 31 ] and rainbow trout intestine [ 26 ], whereas the presence of a mixed population of epithelial and non-epithelial cells has never been explored before in intestinal cell lines of either fish or mammals. In vivo, the lgr5 + cells are located in the stromal tissue and are involved in the maintenance of tissue homeostasis by producing growth factors, cytokines, and proteins of the extracellular matrix [ 29 ]. In vitro, the presence of myofibroblasts enables the successful maintenance of human intestinal epithelial cell formation and proliferation beyond just a few days, even without the presence of supportive growth factors [ 38 ].…”

Organoids and Colorectal Cancer

“…This breakthrough has given rise to several possibilities for customizing microfluidic devices such as co-cultures of organoids with stromal cells. Thus, Verhulsel and colleagues developed a 3D biomimetic scaffold made entirely of collagen treated with threose that cross-links the collagen and thus maintains the desired topography allowing the co-culture of fibroblasts and intestinal organoids [ 182 ]. Colon organoid co-cultures within a self-assembled vascular network have recently been successfully achieved, leading to better cell/organoid growth under constant perfusion than with a conventional static condition [ 183 ].…”

Mechanical Strain-Enabled Reconstitution of Dynamic Environment in Organ-on-a-Chip Platforms: A Review

“…In addition to widely reported desirable effects caused by shear or luminal flow in gut-on-a-chip systems [ 71 , 72 , 73 ], it has been demonstrated that a cyclic strain with physiologically relevant frequency and magnitude can also enhance proliferation and differentiation of human intestinal epithelial cells [ 74 ]. By utilizing external mechanical actuators, the muscle contractions or peristaltic flow can be accurately mimicked in gut-on-a-chip systems.…”