Human cell models to study small intestinal functions: Recapitulation of the crypt‐villus axis

Abstract

The intestinal epithelium is continuously and rapidly renewed by a process involving cell generation, migration, and differentiation, from the stem cell population located at the bottom of the crypt to the extrusion of the terminally differentiated cells at the tip of the villus. Because of the lack of normal human intestinal cell models, most of our knowledge about the regulation of human intestinal cell functions has been derived from studies conducted on cell cultures generated from experimental animals and human colon cancers. However, important advances have been achieved over recent years in the generation of normal human intestinal cell models. These models include (a) intestinal cell lines with typical crypt cell proliferative noncommitted characteristics, (b) conditionally immortalized intestinal cell lines that can be induced to differentiate, and (c) primary cultures of differentiated villuslike cells that can be maintained in culture for up to 10 days. Each of these models should help in the investigation of the specific aspects of human intestinal function and regulation. Furthermore, taken together, these models provide an integrated system that allows an in vitro recapitulation of the entire crypt-villus axis of the normal human small intestine.

Culture of human intestinal epithelial cell using the dissociating enzyme thermolysin and endothelin-3

“…As a crypt cell marker, the detection of the MIM-1/39 antigen provided strong evidence that serially passaged HIEC still retained their epithelial character, a feature that was absent in Caco-2 and HIM cells (21,23). This may also indicate that HIEC had the characteristics of crypt cells (31). The absence of MGA expression in HIEC was also demonstrated in previous experiments (6); this was expected since the enzymatic complex has been reported to be absent from the human fetal intestine (32).…”

Proinflammatory cytokines TNF-α and IFN-γ alter laminin expression under an apoptosis-independent mechanism in human intestinal epithelial cells

“…The normal embryonic human intestinal epithelial cell line HIEC-6 was generated as described previously (39). These cells, which express a number of intestinal cell markers but no villus cell markers, are representative of the intestinal crypt epithelium (37). HIEC cells were routinely grown in OPTI-MEM (GIBCO-BRL, Rockville, MD) supplemented with 20 mM HEPES, 10 mM GlutaMax (both from GIBCO-BRL), 5 ng/ml EGF (Sigma, St. Louis, MO), and 4% FBS (CELLect Gold, ICN Biochemical, Aurora, OH).…”

“…To further investigate the epithelial regulation of laminin expression in the crypt BM, we used the well-characterized normal human intestinal epithelial cell line HIEC, representative of the crypt epithelium (37,39).…”

“…Considering that changes in extracellular matrix production in CD are mainly observed in the lower two-thirds of the crypt, we have tested the cytokine effects on laminin production by using the well-characterized human intestinal epithelial crypt cell line (HIEC) (39) as an experimental cell model representative of the proliferative compartment of the human intestinal lower crypt (37).…”

Human Prenatal Small Intestine Cells as a Valuable Source of Stem Cells and Epithelial Cells: Phenotypic and Functional Characterization

“…2 Small intestine epithelial cells are potentially a suitable model for researching inflammatory bowel disease, colitis, intestinal cancer, intestinal infections and more importantly, toxicity and drug discovery. Although recent research has described a variety of culture methods, [3][4][5][6][7] no simplified long-term culture system has been established from human prenatal intestinal tissue that maintains basic crypt-villus physiology and metabolic functionality in the differentiated state. The majority of studies are not performed on primary intestinal epithelial cells that have been derived directly from human intestinal tissue.…”