Characterization of conditions for the primary culture of human small intestinal epithelial cells

Aldhous, Marian C.; Shmakov, Andrei N.; Bode, John; Ghosh, Subrata

doi:10.1046/j.1365-2249.2001.01522.x

Cited by 28 publications

(26 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…(4) DMEM/F12 mix (Sigma-Aldrich): like DMEM 4.5, the DMEM/F12 mix has also been used successfully to culture epithelial cells [Aldhous et al, 2001;Schierack et al, 2006;Bridger et al, 2007;Koh et al, 2008]. Thus, we tested whether it would be more suitable for culturing colonic epithelial cells than DMEM 4.5.…”

Section: Epithelial Cell Isolation and Culturementioning

confidence: 99%

Establishment and Characterization of Porcine Colonic Epithelial Cells Grown in Primary Culture

Petto

Lesko

Gäbel

et al. 2011

Cells Tissues Organs

View full text Add to dashboard Cite

Background: Primary cultures of epithelial cells are suitable models for studying epithelial function and, in particular, the regulation of epithelial tightness in vitro. The aim of our study was to develop a protocol for the isolation and culture of porcine colonic epithelial cells and to establish transepithelial electrical resistance (TEER) as a functional parameter for epithelial tightness. Methods: Epithelial cells were obtained from the proximal colon of piglets by enzymatic dispase digestion. Cells were cultured on collagen-coated membrane supports for 21 days. The epithelial origin of the cells was shown by immunohistochemical detection of cytokeratin and zonula occludens protein 1 (ZO-1). Scanning electron microscopy, transmission electron microscopy and confocal microscopy were used for further morphological characterization. The integrity and tightness of the artificial epithelium were determined by measuring TEER. Results: The cultured epithelial cells were immunoreactive for cytokeratin and ZO-1. They showed dense microvilli on their apical membranes and expression of Na⁺/K⁺-ATPase on their basolateral membranes. Adjacent cells were connected by tight junctions. We observed TEER to continuously increase up to 870 ± 38 Ω·cm² during the culture period. TEER correlated with the amount of epithelial cells expressing ZO-1. Conclusions: The properties of primary cultured epithelial cells resemble the structural properties of polarized colonic epithelium in vivo. Measurement of TEER seems to be suitable for studying epithelial tightness in vitro. We suggest that these primary epithelial cultures be used to investigate the regulation of the epithelial barrier function.

show abstract

Section: Epithelial Cell Isolation and Culturementioning

confidence: 99%

Establishment and Characterization of Porcine Colonic Epithelial Cells Grown in Primary Culture

Petto

Lesko

Gäbel

et al. 2011

Cells Tissues Organs

View full text Add to dashboard Cite

show abstract

“…The utilization of human primary normal enterocytes was expected to be a promising solution for the aforementioned discrepancy between in vivo and in vitro models. Although several methods have been used to isolate primary enterocytes from the human small intestine (Perreault and Beaulieu, 1998;Aldhous et al, 2001;Grossmann et al, 2003;Chougule et al, 2012), their applications remain limited, which has been attributed to their poor viability and short life span (Aldhous et al, 2001;Grossmann et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

Human Small Intestinal Epithelial Cells Differentiated from Adult Intestinal Stem Cells as a Novel System for Predicting Oral Drug Absorption in Humans

et al. 2014

View full text Add to dashboard Cite

Adult intestinal stem cells (ISCs) possess both a long-term proliferation ability and differentiation capability into enterocytes. As a novel in vitro system for the evaluation of drug absorption, we characterized a human small intestinal epithelial cell (HIEC) monolayer that differentiated from adult ISCs. Continuous proliferation/ differentiation from ISCs consistently conferred the capability of maturation of enterocytes to HIECs over 25 passages. The morphologically matured HIEC monolayer consisted of polarized columnar epithelia with dense microvilli, tight junctions, and desmosomes 8 days after seeding onto culture inserts. Transepithelial electrical resistance across the monolayer was 9-fold lower in HIECs (98.9 V 3 cm 2 ) than in Caco-2 cells (900 V 3 cm 2 ), which indicated that the looseness of the tight junctions in the HIEC monolayer was similar to that in the human small intestine (approximately 40 V 3 cm 2 ). No significant differences were observed in the overall gene expression patterns of the major drug-metabolizing enzymes and transporters between the HIEC and Caco-2 cell monolayers. Furthermore, the functions of P-glycoprotein and breast cancer resistance protein in the HIEC monolayer were confirmed by the vectorial transport of marker substrates and their disappearance in the presence of specific inhibitors. The apparent drug permeability values of paracellularly transported compounds (fluorescein isothiocyanate-dextran 4000, atenolol, and terbutaline) and nucleoside transporter substrates (didanosine, ribavirin, and doxifluridine) in the HIEC monolayer were markedly higher than those of Caco-2 cells, whereas transcellularly transported drugs (pindolol and midazolam) were equally well permeated. In conclusion, the HIEC monolayer can serve as a novel and superior alternative to the conventional Caco-2 cell monolayer for predicting oral absorption in humans.

show abstract

“…During past decades, primary intestinal cell cultures have been obtained from the small intestine (Perreault and Beaulieu, 1998;Aldhous et al, 2001) or from the colon (Baten et al, 1992;Fonti et al, 1994;Deveney et al, 1996;Grossmann et al, 2003;Mohammadpour, 2005), but a rapid loss of the differentiated charac-teristics of cells was observed. As a consequence, these primary cell models could be used only for short-term toxicity studies.…”

Section: Introductionmentioning

confidence: 99%

Gene Expression Profiling of Systems Involved in the Metabolism and the Disposition of Xenobiotics: Comparison between Human Intestinal Biopsy Samples and Colon Cell Lines

Bourgine¹,

Billaut-Laden²,

Happillon³

et al. 2012

Drug Metab Dispos

View full text Add to dashboard Cite

ABSTRACT:Intestinal cell lines are used as in vitro models for pharmacological and toxicological studies. However, a general report of the gene expression spectrum of proteins that are involved in the metabolism and the disposition of xenobiotics in these in vitro systems is not currently available. To fill this information gap, we systematically characterized the expression profile of 377 genes encoding xenobiotic-metabolizing enzymes, transporters, and nuclear receptors and transcription factors in intestinal mucosa (ileum, ascending colon, transverse colon, descending colon, and rectum) from five healthy subjects and in five commonly used intestinal cell lines (Caco-2, C2BBe1, HT29, T84, and FHC).

show abstract

Characterization of conditions for the primary culture of human small intestinal epithelial cells

Cited by 28 publications

References 26 publications

Establishment and Characterization of Porcine Colonic Epithelial Cells Grown in Primary Culture

Establishment and Characterization of Porcine Colonic Epithelial Cells Grown in Primary Culture

Human Small Intestinal Epithelial Cells Differentiated from Adult Intestinal Stem Cells as a Novel System for Predicting Oral Drug Absorption in Humans

Gene Expression Profiling of Systems Involved in the Metabolism and the Disposition of Xenobiotics: Comparison between Human Intestinal Biopsy Samples and Colon Cell Lines

Contact Info

Product

Resources

About