Long-Term Culture of Intestinal Cell Progenitors: An Overview of Their Development, Application, and Associated Technologies

Abstract

Purpose of ReviewLong-term culture of adult progenitor cells in 3D is a recently emerging technology that inhabits the space between 2D cell lines and organ slice culture.Recent FindingsAdaptations to defined media components in the wake of advances in ES and iPS cell culture has led to the identification of conditions that maintained intestinal cell progenitors in culture. These conditions retain cellular heterogeneity of the normal or tumour tissue, and the cultures have been shown to be genetically stable, such that substantial biobanks are being created from patient derived material. This coupled with advances in analytical tools has generated a field, characterized by the term “organoid culture”, that has huge potential for advancing drug discovery, regenerative medicine, and furthering the understanding of fundamental intestinal biology.SummaryIn this review, we describe the approaches available for the long-term culture of intestinal cells from normal and diseased tissue, the current challenges, and how the technology is likely to develop further.

Organoid-based Models to Study the Role of Host-microbiota Interactions in IBD

“…This aspect is very important for IBD, in which defects in several epithelial cell type functions have been observed. 18 However, for other questions, more ‘complex’ in vitro research models must include other cell types known to be essential in intestinal physiology and pathophysiology, 54 , 88 including a functional immune system, enteric nerves, or nutrients providing the mesenchymal niche. 89 Co-culture with immune cells [macrophages, 83 neutrophils, 71 T lymphocytes, 90 and intraepithelial lymphocytes, 91 ] as well as fibroblasts, 92 adipocytes, 93 and enteric nervous system cells, 94 have been key to gain knowledge on the role of these cell types in host-microbial interactions in IBD.…”

Organoids, organs-on-chips and other systems, and microbiota

“…• Near-physiological model for epithelial tissue containing heterogeneous cell populationsmaintaining integrity of the in vivo structure [49].…”

“…Long-term organoid culturing has been a major breakthrough in research and is now an extensively used tool for studying both basic and clinical biology [49][50][51]. Advancement in technology has enabled culturing of the stomach [52][53][54], pancreas [55][56][57], liver [58], prostate [59,60], oesophagus [46,61], gall bladder [62,63], and taste buds [64].…”

“…It is not possible to exert peristalsis-like motions (which is required in vivo to sustain intestinal differentiation and confine microbial overgrowth [14]) to the cells cultured in Transwell or organoid culture. And the reductionist approach of organoid culture limits the full potential of this method as epithelial cells grow without their native microenvironment, and the true contributions from the stroma, extracellular matrix (ECM), oxygen, vasculature, immune system, and the microbiome cannot be truly assessed [49,79]. This ultimately impedes the understanding of the precise molecular mechanisms involved in transition from healthy to disease states and limits the potential efficacy of interventions when translated to clinic.…”