Human intestinal goblet cells in monolayer culture: Characterization of a mucus-secreting subclone derived from the HT29 colon adenocarcinoma cell line

Phillips, Thomas E.; Huet, Christian; Bilbo, Patrick; Podolsky, Daniel K.; Louvard, Daniel; Neutra, Marian R.

doi:10.1016/0016-5085(88)90678-6

Cited by 92 publications

(55 citation statements)

References 26 publications

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“…For experiments, LLC-PK1 cells were used 4 to 6 days after confluence when domes had fully developed. For experiments with various concentrations of glucose or other monosaccharides, glucose-free medium (32) containing dialyzed fetal bovine serum was prepared.…”

Section: Methodsmentioning

confidence: 99%

Regulation of glucose transporters in LLC-PK1 cells: effects of D-glucose and monosaccharides.

1990

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Regulation of D-glucose transport in the porcine kidney epithelial cell line LLC-PK, was examined. To identify the sodium-coupled glucose transporter (SGLT), we cloned and sequenced several partial cDNAs homologous to SGLT1 from rabbit small intestine (M. A. Hediger, M. J. Coady, T. S. Ikeda, and E. M. Wright, Nature (London) 330: [379][380][381] 1987). The extensive homology of the two sequences leads us to suggest that the high-affinity SGLT expressed by LLC-PK1 cells is SGLT1. SGLT1 mRNA levels were highest when the D-glucose concentration in the culture medium was 5 to 10 mM. Addition of D-mannose or D-fructose, but not D-galactose, in the presence of 5 mM D-glucose suppressed SGLT1 mRNA levels. SGLT1 activity, measured by methyl a-D-glucopyranoside uptake, paralleled message levels except in cultures containing D-galactose. Therefore, SGLT1 gene expression may respond either to the cellular energy status or to the concentration of a hexose metabolite(s). By isolating several cDNAs homologous to rat GLUT-1, we identified the facilitated glucose transporter in LLC-PK, cells as the erythroid/brain type GLUT-1. High-stringency hybridization of a single mRNA transcript to the rat GLUT-1 cDNA probe and failure to observe additional transcripts hybridizing either to GLUT-1 or to GLUT-2 probes at low stringency provide evidence that GLUT-1 is the major facilitated glucose transporter in this cell line. LLC-PK, GLUT-1 mRNAs were highest at medium nglucose concentrations of s2 mM. D-Fructose, D-mannose, and to a lesser extent n-galactose all suppressed GLUT-1 mRNA levels. Since the pattern of SGLT1 and GLUT-1 expression differed, particularly in low Dglucose or in the presence of D-galactose, we suggest that the two transporters are regulated independently.

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Section: Methodsmentioning

confidence: 99%

Regulation of glucose transporters in LLC-PK1 cells: effects of D-glucose and monosaccharides.

1990

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“…A second mucus-secreting, clonal derivative (HT29-SB) has also been established (59). Growing the parental HT-29 cell line in the absence of glucose results in the selection of homogeneous columnar cells with the typical goblet cell morphology, which have been designated the HT29-18N2 clone (41,60). Adaptation of the parental HT-29 cell line to lethal concentrations of methotrexate (MTX) (38) and 5-fluorouracil (FU) (61) has been shown to result in the emergence of subpopulations of cells that are all stably committed to differentiation (38,61).…”

Section: Differentiated Mucin-secreting Ht-29 Cell Subpopulations Andmentioning

confidence: 99%

Pathogenesis of Human Enterovirulent Bacteria: Lessons from Cultured, Fully Differentiated Human Colon Cancer Cell Lines

Moal

Servin

2013

Microbiol Mol Biol Rev

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SUMMARY Hosts are protected from attack by potentially harmful enteric microorganisms, viruses, and parasites by the polarized fully differentiated epithelial cells that make up the epithelium, providing a physical and functional barrier. Enterovirulent bacteria interact with the epithelial polarized cells lining the intestinal barrier, and some invade the cells. A better understanding of the cross talk between enterovirulent bacteria and the polarized intestinal cells has resulted in the identification of essential enterovirulent bacterial structures and virulence gene products playing pivotal roles in pathogenesis. Cultured animal cell lines and cultured human nonintestinal, undifferentiated epithelial cells have been extensively used for understanding the mechanisms by which some human enterovirulent bacteria induce intestinal disorders. Human colon carcinoma cell lines which are able to express in culture the functional and structural characteristics of mature enterocytes and goblet cells have been established, mimicking structurally and functionally an intestinal epithelial barrier. Moreover, Caco-2-derived M-like cells have been established, mimicking the bacterial capture property of M cells of Peyer's patches. This review intends to analyze the cellular and molecular mechanisms of pathogenesis of human enterovirulent bacteria observed in infected cultured human colon carcinoma enterocyte-like HT-29 subpopulations, enterocyte-like Caco-2 and clone cells, the colonic T84 cell line, HT-29 mucus-secreting cell subpopulations, and Caco-2-derived M-like cells, including cell association, cell entry, intracellular lifestyle, structural lesions at the brush border, functional lesions in enterocytes and goblet cells, functional and structural lesions at the junctional domain, and host cellular defense responses.

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“…extending from C72) were isolated from human colonic libraries. Thus, we constructed a cDNA library of human N2 cell line, which can differentiate into goblet cells and express Fc␥BP (25,26). Using a 5Ј-terminal probe derived from the C72 clone, we obtained a clone, NZ4, containing the 5Ј-terminal ATG.…”

Section: Cloning and Sequencing Of Full-length Cdna For Humanmentioning

confidence: 99%

Human IgGFc Binding Protein (FcγBP) in Colonic Epithelial Cells Exhibits Mucin-like Structure

Harada¹,

Iijima²,

Kobayashi³

et al. 1997

Journal of Biological Chemistry

104

127

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Human intestinal goblet cells in monolayer culture: Characterization of a mucus-secreting subclone derived from the HT29 colon adenocarcinoma cell line

Cited by 92 publications

References 26 publications

Regulation of glucose transporters in LLC-PK1 cells: effects of D-glucose and monosaccharides.

Regulation of glucose transporters in LLC-PK1 cells: effects of D-glucose and monosaccharides.

Pathogenesis of Human Enterovirulent Bacteria: Lessons from Cultured, Fully Differentiated Human Colon Cancer Cell Lines

Human IgGFc Binding Protein (FcγBP) in Colonic Epithelial Cells Exhibits Mucin-like Structure

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