Differentiation of rat small intestinal epithelial cells by extracellular matrix

1988 ◽  
Vol 254 (3) ◽  
pp. G355-G360 ◽  
Author(s):  
K. M. Carroll ◽  
T. T. Wong ◽  
D. L. Drabik ◽  
E. B. Chang
Keyword(s):  
Extracellular Matrix ◽  
Basement Membrane ◽  
Glucose Absorption ◽  
Cell Maturation ◽  
Intestinal Cell ◽  
Synthetic Activity ◽  
Intestinal Epithelial ◽  
Intestinal Cell Line ◽  
Small Intestinal

The role of extracellular matrix as a determinant of intestinal cell maturation was explored by growing a normal, but immature, rat small intestinal cell line (IEC-6) on basement membrane extract from Engelbreth-Holm-Swarm (EHS) sarcoma cells (ECM). Grown on plastic or glass, these cells are relatively immature and proliferate rapidly. In contrast, cells on ECM attached more rapidly, stopped proliferating, and rapidly organized into multicellular complex structures. Ultrastructurally, cells grown on ECM displayed significantly more mitochondria, rough endoplasmic reticulum, apical microvilli, and complex golgi apparatus, consistent with greater maturity and synthetic activity. By indirect immunofluorescence, sucrase, alkaline phosphatase, and cellular apolipoprotein B were present in cells grown on ECM only. In contrast to cells grown on glass, these cells also demonstrated Na-dependent glucose absorption, a function unique to mature villus cells (7). We conclude that the basement membrane may be a key determinant of intestinal epithelial cell maturation. The development of a mature villuslike intestinal cell in vitro may have wide application for future studies of induction and regulation of intestinal maturation and function.

scholarly journals Primary Murine Small Intestinal Epithelial Cells, Maintained in Long-Term Culture, Are Susceptible to Rotavirus Infection

2000 ◽  
Vol 74 (12) ◽  
pp. 5597-5603 ◽  
Author(s):  
Kristine K. Macartney ◽  
Daniel C. Baumgart ◽  
Simon R. Carding ◽  
Jeffery O. Brubaker ◽  
Paul A. Offit
Keyword(s):  
Epithelial Cells ◽  
Mhc Class Ii ◽  
Intestinal Epithelial Cells ◽  
Intestinal Cell ◽  
Intestinal Epithelial ◽  
Long Term Culture ◽  
Intestinal Cell Line ◽  
Small Intestinal

ABSTRACT We describe a method for long-term culture of primary small intestinal epithelial cells (IEC) from suckling mice. IEC were digested from intestinal fragments as small intact units of epithelium (organoids) by using collagenase and dispase. IEC proliferated from organoids on a basement-membrane-coated culture surface and remained viable for 3 weeks. Cultured IEC had the morphologic and functional characteristics of immature enterocytes, notably sustained expression of cytokeratin and alkaline phosphatase. Few mesenchymal cells were present in the IEC cultures. IEC were also cultured from adult BALB/c mice and expressed major histocompatibility complex (MHC) class II antigens for at least 48 h in vitro. Primary IEC supported the growth of rhesus rotavirus (RRV) to a greater extent than a murine small intestinal cell line, m-ICcl2. Cell-culture-adapted murine rotavirus strain EDIM infected primary IEC and m-ICcl2 cells to a lesser extent than RRV. Wild-type EDIM did not infect either cell type. Long-term culture of primary murine small intestinal epithelial cells provides a method to study (i) virus-cell interactions, (ii) the capacity of IEC to act as antigen-presenting cells using a wide variety of MHC haplotypes, and (iii) IEC biology.

scholarly journals Cellular Differentiation of Non-Transformed Intestinal Epithelial Cells Is Regulated by Lactobacillus rhamnosus and L. casei Strains

2018 ◽  
pp. 261-273
Author(s):  
J. KOLINSKA ◽  
M. ZAKOSTELECKA ◽  
Z. ZEMANOVA ◽  
V. LISA ◽  
J. GOLIAS ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Cellular Differentiation ◽  
Detection System ◽  
Lactobacillus Rhamnosus ◽  
Immunofluorescent Staining ◽  
Intestinal Cell ◽  
Intestinal Epithelial ◽  
Caspase 1 ◽  
Intestinal Cell Line

The aim of this study was to characterize an in vitro modulating effect of three commensal Lactobacillus strains on cellular differentiation of non-transformed crypt-like rat small intestinal cell line IEC-18. IEC-18 was grown on extracellular matrix, with or without presence of Lactobacillus strains. Gene expression of IEC-18 bacterial detection system – such as Toll-like receptors TLR-2, TLR-4, signal adapter MyD88, cytoplasmic NOD2 receptor, inflammatory cytokines IL-18, IL-1β, chemokine IL-8 and enzyme caspase-1 – was evaluated using real-time PCR. Expression and localization of TLR-2, TLR-4, IL-18 and caspase-1 proteins was demonstrated by Western blotting and immunofluorescent staining. Secretion of IL-18 to apical and basolateral surfaces was assayed by ELISA. Our results suggested that L. casei LOCK0919 accelerated differentiation of IEC-18 by stimulating TLR-2, TLR-4, MyD88, IL-18, caspase-1 mRNAs and proteins. L. casei LOCK0919 increased expression and transfer of villin and β-catenin from cytoplasm to cell membrane. Presence of L. rhamnosus LOCK0900 resulted in detachment of IEC-18 layer from extracellular matrix leading to induction of IL-1β, of TLR-2 and IL-8 mRNAs and stimulation of MyD88, caspase-1 and cytosolic receptor NOD2 mRNAs. L. rhamnosus LOCK0908 was not recognized by TLR-2 or TLR-4 receptors. Lactobacilli-IEC-18 crosstalk enhanced immune and barrier mucosal functions.

scholarly journals Epithelioid cell cultures from rat small intestine. Characterization by morphologic and immunologic criteria.

1979 ◽  
Vol 80 (2) ◽  
pp. 248-265 ◽  
Author(s):  
A Quaroni ◽  
J Wands ◽  
R L Trelstad ◽  
K J Isselbacher
Keyword(s):  
Basement Membrane ◽  
Amorphous Material ◽  
Population Doubling ◽  
Antigenic Determinants ◽  
Intestinal Epithelial ◽  
Epithelioid Cells ◽  
Intestinal Crypt ◽  
Ultrastructural Studies ◽  
Small Intestinal

Rat small intestinal epithelial cell lines have been established in vitro and subcultured serially for periods up to 6 mo. These cells have an epithelioid morphology, grow as monolayers of closely opposed polygonal cells, and during the logarithmic phase of growth have a population doubling time of 19--22 h. Ultrastructural studies revealed the presence of microvilli, tight junctions, an extensive Golgi complex, and the presence of extracellular amorphous material similar in appearance to isolated basement membrane. These cells exhibit a number of features characteristic of normal cells in culture; namely, a normal rat diploid karyotype, strong density inhibition of growth, lack of growth in soft agar, and a low plating efficiency when seeded at low density. They did not produce tumors when injected in syngeneic animals. Immunochemical studies were performed to determine their origin using antisera prepared against rat small intestinal crypt cell plasma membrane, brush border membrane of villus cells and isolated sucrase-isomaltase complex. Antigenic determinants specific for small intestinal epithelial (crypt and villus) cells were demonstrated on the surface of the epithelioid cells, but they lacked immunological determinants specific for differentiated villus cells. An antiserum specifically staining extracellular material surrounding the cells cultured in vitro demonstrated cross-reactivity to basement membrane in rat intestinal frozen sections. It is concluded that the cultured epithelioid cells have features of undifferentiated small intestinal crypt cells.

scholarly journals Traditional Herbal Medicine, Rikkunshito, Induces HSP60 and Enhances Cytoprotection of Small Intestinal Mucosal Cells as a Nontoxic Chaperone Inducer

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Kumiko Tamaki ◽  
Michiro Otaka ◽  
Tomoyoshi Shibuya ◽  
Naoto Sakamoto ◽  
Soh Yamamoto ◽  
...  
Keyword(s):  
Herbal Medicine ◽  
Immunoblot Analysis ◽  
Intestinal Cell ◽  
Traditional Herbal Medicine ◽  
Intestinal Cell Line ◽  
Mucosal Cells ◽  
Hsp60 Expression ◽  
Small Intestinal ◽  
Intestinal Ulcers

Increasing incidence of small intestinal ulcers associated with nonsteroidal anti-inflammatory drugs (NSAIDs) has become a topic with recent advances of endoscopic technology. However, the pathogenesis and therapy are not fully understood. The aim of this study is to examine the effect of Rikkunshito (TJ-43), a traditional herbal medicine, on expression of HSP60 and cytoprotective ability in small intestinal cell line (IEC-6). Effect of TJ-43 on HSP60 expression in IEC-6 cells was evaluated by immunoblot analysis. The effect of TJ-43 on cytoprotective abilities of IEC-6 cells against hydrogen peroxide or indomethacin was studied by MTT assay, LDH-release assay, caspase-8 activity, and TUNEL. HSP60 was significantly induced by TJ-43. Cell necrosis and apoptosis were significantly suppressed in IEC-6 cells pretreated by TJ-43 with overexpression of HSP60. Our results suggested that HSP60 induced by TJ-43 might play an important role in protecting small intestinal epithelial cells from apoptosis and necrosis in vitro.

Helicobacter pylori Stimulates DNA Synthesis in a Small Intestinal Cell Line in vitro

Digestion ◽  
1998 ◽  
Vol 59 (1) ◽  
pp. 33-39 ◽  
Author(s):  
Johanna Brännström ◽  
Kristina Zachrisson ◽  
Kristina Hultén ◽  
Lars Engstrand ◽  
Andrés Uribe
Keyword(s):  
Helicobacter Pylori ◽  
Dna Synthesis ◽  
Cell Line ◽  
Intestinal Cell ◽  
Intestinal Cell Line ◽  
Small Intestinal

scholarly journals Monocyte Differentiation into Destructive Macrophages on In Vitro Administration of Gingival Crevicular Fluid from Periodontitis Patients

2021 ◽  
Vol 11 (6) ◽  
pp. 555
Author(s):  
Hammam Ibrahim Fageeh ◽  
Hytham N. Fageeh ◽  
Shankargouda Patil
Keyword(s):  
Stem Cells ◽  
Extracellular Matrix ◽  
Stromal Cells ◽  
Pathogenic Bacteria ◽  
Gingival Crevicular Fluid ◽  
Gingival Tissue ◽  
Synthetic Activity ◽  
Periodontal Destruction ◽  
Crevicular Fluid

Background: Periodontitis is an inflammatory condition of the tooth-supporting structures initiated and perpetuated by pathogenic bacteria present in the dental plaque biofilm. In periodontitis, immune cells infiltrate the periodontium to prevent bacterial insult. Macrophages derived from monocytes play an important role in antigen presentation to lymphocytes. However, they are also implicated in causing periodontal destruction and bystander damage to the host tissues. Objectives: The objective of the present study was to quantify the cytokine profile of gingival crevicular fluid (GCF) samples obtained from patients with periodontitis. The study further aimed to assess if GCF of periodontitis patients could convert CD14+ monocytes into macrophages of destructive phenotype in an in vitro setting. The secondary objectives of the study were to assess if macrophages that resulted from GCF treatment of monocytes could affect the synthetic properties, stemness, expression of extracellular matrix proteins, adhesion molecules expressed by gingival stem cells, gingival mesenchymal stromal cells, and osteoblasts. Methods: GCF, blood, and gingival tissue samples were obtained from periodontitis subjects and healthy individuals based on specific protocols. Cytokine profiles of the GCF samples were analyzed. CD14+ monocytes were isolated from whole blood, cultured, and treated with the GCF of periodontitis patients to observe if they differentiated into macrophages. Further, the macrophages were assessed for a phenotype by surface marker analysis and cytokine assays. These macrophages were co-cultured with gingival stem cells, epithelial, stromal cells, and osteoblasts to assess the effects of the macrophages on the synthetic activity of the cells. Results: The GCF samples of periodontitis patients had significantly higher levels of IFN gamma, M-CSF, and GM-CSF. Administration of the GCF samples to CD14+ monocytes resulted in their conversion to macrophages that tested positive for CD80, CD86, and CD206. These macrophages produced increased levels of IL-1β, TNF-α, and IL-6. Co-culture of the macrophages with gingival stem cells, epithelial cells, and stromal cells resulted in increased cytotoxicity and apoptotic rates to the gingival cells. A reduced expression of markers related to stemness, extracellular matrix, and adhesion namely OCT4, NANOG, KRT5, POSTN, COL3A1, CDH1, and CDH3 were seen. The macrophages profoundly affected the production of mineralized nodules by osteoblasts and significantly reduced the expression of COL1A1, OSX, and OCN genes. Conclusion: In periodontitis patients, blood-derived monocytes transform into macrophages of a destructive phenotype due to the characteristic cytokine environment of their GCF. Further, the macrophages affect the genotype and phenotype of the resident cells of the periodontium, aggravate periodontal destruction, as well as jeopardize periodontal healing and resolution of inflammation.

scholarly journals “Masato de Yuca” and “Chicha de Siete Semillas” Two Traditional Vegetable Fermented Beverages from Peru as Source for the Isolation of Potential Probiotic Bacteria

2021 ◽  
Author(s):  
Teresa D. Rebaza-Cardenas ◽  
Kenneth Silva-Cajaleón ◽  
Carlos Sabater ◽  
Susana Delgado ◽  
Nilda D. Montes-Villanueva ◽  
...  
Keyword(s):  
Gastrointestinal Transit ◽  
Probiotic Strain ◽  
In Vitro Digestion ◽  
Intestinal Cell ◽  
Adhesion Ability ◽  
Intestinal Cell Line ◽  
Carbohydrate Fermentation ◽  
Traditional Vegetable ◽  
Genetic Profiles

AbstractIn this work, two Peruvian beverages “Masato de Yuca,” typical of the Amazonian communities made from cassava (Manihot esculenta), and “Chicha de Siete Semillas,” made from different cereal, pseudo-cereal, and legume flours, were explored for the isolation of lactic acid bacteria after obtaining the permission of local authorities following Nagoya protocol. From an initial number of 33 isolates, 16 strains with different RAPD- and REP-PCR genetic profiles were obtained. In Chicha, all strains were Lactiplantibacillus plantarum (formerly Lactobacillus plantarum), whereas in Masato, in addition to this species, Limosilactobacillus fermentum (formerly Lactobacillus fermentum), Pediococcus acidilactici, and Weissella confusa were also identified. Correlation analysis carried out with their carbohydrate fermentation patterns and enzymatic profiles allowed a clustering of the lactobacilli separated from the other genera. Finally, the 16 strains were submitted to a static in vitro digestion (INFOGEST model) that simulated the gastrointestinal transit. Besides, their ability to adhere to the human epithelial intestinal cell line HT29 was also determined. Following both procedures, the best probiotic candidate was Lac. plantarum Ch13, a robust strain able to better face the challenging conditions of the gastrointestinal tract and showing higher adhesion ability to the intestinal epithelium in comparison with the commercial probiotic strain 299v. In order to characterize its benefit for human health, this Ch13 strain will be deeply studied in further works.

scholarly journals Gluten-induced RNA methylation changes regulate intestinal inflammation via allele-specific XPO1 translation in epithelial cells

Gut ◽  
2021 ◽  
pp. gutjnl-2020-322566
Author(s):  
Ane Olazagoitia-Garmendia ◽  
Linda Zhang ◽  
Paula Mera ◽  
Julie K Godbout ◽  
Maialen Sebastian-DelaCruz ◽  
...  
Keyword(s):  
Intestinal Inflammation ◽  
Autoimmune Disorder ◽  
Intestinal Cell ◽  
In Vivo Models ◽  
Knock Out ◽  
New Therapeutic Approaches ◽  
Intestinal Cell Line ◽  
Environment Characteristic

ObjectivesCoeliac disease (CD) is a complex autoimmune disorder that develops in genetically susceptible individuals. Dietary gluten triggers an immune response for which the only available treatment so far is a strict, lifelong gluten free diet. Human leucocyte antigen (HLA) genes and several non-HLA regions have been associated with the genetic susceptibility to CD, but their role in the pathogenesis of the disease is still essentially unknown, making it complicated to develop much needed non-dietary treatments. Here, we describe the functional involvement of a CD-associated single-nucleotide polymorphism (SNP) located in the 5’UTR of XPO1 in the inflammatory environment characteristic of the coeliac intestinal epithelium.DesignThe function of the CD-associated SNP was investigated using an intestinal cell line heterozygous for the SNP, N6-methyladenosine (m6A)-related knock-out and HLA-DQ2 mice, and human samples from patients with CD.ResultsIndividuals harbouring the risk allele had higher m6A methylation in the 5’UTR of XPO1 RNA, rendering greater XPO1 protein amounts that led to downstream nuclear factor kappa B (NFkB) activity and subsequent inflammation. Furthermore, gluten exposure increased overall m6A methylation in humans as well as in in vitro and in vivo models.ConclusionWe identify a novel m6A-XPO1-NFkB pathway that is activated in CD patients. The findings will prompt the development of new therapeutic approaches directed at m6A proteins and XPO1, a target under evaluation for the treatment of intestinal disorders.

scholarly journals Effect of Helicobacter pylori on Polymorphonuclear Leukocyte Migration across Polarized T84 Epithelial Cell Monolayers: Role of Vacuolating Toxin VacA and cagPathogenicity Island

2000 ◽  
Vol 68 (9) ◽  
pp. 5225-5233 ◽  
Author(s):  
Véronique Hofman ◽  
Vittorio Ricci ◽  
Antoine Galmiche ◽  
Patrick Brest ◽  
Patrick Auberger ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Polymorphonuclear Leukocyte ◽  
Broth Culture ◽  
Intestinal Cell ◽  
T84 Cells ◽  
Vacuolating Cytotoxin ◽  
Intestinal Cell Line ◽  
H Pylori

ABSTRACT Helicobacter pylori infection can induce polymorphonuclear leukocyte (PMNL) infiltration of the gastric mucosa, which characterizes acute chronic gastritis. The mechanisms underlying this process are poorly documented. The lack of an in vitro model has considerably impaired the study of transepithelial migration of PMNL induced by H. pylori. In the present work, we used confluent polarized monolayers of the human intestinal cell line T84 grown on permeable filters to analyze the epithelial PMNL response induced by broth culture filtrates (BCFs) and bacterial suspensions from different strains of H. pylori. We have evaluated the role of the vacuolating cytotoxin VacA and of the cagpathogenicity island (PAI) of H. pylori in PMNL migration via their effects on T84 epithelial cells. We noted no difference in the rates of PMNL transepithelial migration after epithelial preincubation with bacterial suspensions or with BCFs of VacA-negative or VacA-positive H. pylori strains. In contrast, PMNL transepithelial migration was induced after incubation of the T84 cells with cag PAI-positive and cagE-positiveH. pylori strains. Finally, PMNL migration was correlated with a basolateral secretion of interleukin-8 by T84 cells, thus creating a subepithelial chemotactic gradient for PMNL. These data provide evidence that the vacuolating cytotoxin VacA is not involved in PMNL transepithelial migration and that the cag PAI, with a pivotal role for the cagE gene, provokes a transcellular signal across T84 monolayers, inducing a subepithelial PMNL response.

A thermal injury-induced circulating factor(s) compromises intestinal cell morphology, proliferation, and migration

1999 ◽  
Vol 277 (1) ◽  
pp. G175-G182 ◽  
Author(s):  
Maryam Varedi ◽  
George H. Greeley ◽  
David N. Herndon ◽  
Ella W. Englander
Keyword(s):  
Epithelial Cells ◽  
Thermal Injury ◽  
Intestinal Epithelial Cells ◽  
Intestinal Cell ◽  
Intestinal Epithelial ◽  
Rat Serum ◽  
In Vitro Effects ◽  
Mucosal Morphology ◽  
And Migration

The effects of a 60% body surface area thermal injury in rats on the morphology and proliferation of the epithelium of the small intestine and the in vitro effects of serum collected from scalded rats on intestinal epithelial cells were investigated. Scald injury caused significant reductions in duodenal villus width and crypt dimensions, villus enterocytes changed in shape from columnar to cuboidal, and the number of goblet cells decreased. The proportion of bromodeoxyuridine-labeled S phase cells in crypts was also diminished. In vitro, incubation of intestinal epithelial cells (IEC-6) with scalded rat serum (SRS) collected at either 12 or 24 h after injury caused a disruption in the integrity of the confluent culture and induced the appearance of large denuded areas. SRS also decreased DNA synthesis and delayed wound closure in an in vitro wound-healing model. The thermal injury-induced changes in intestinal mucosal morphology and epithelial cell growth characteristics described in this study may underlie, in part, the mechanism(s) involved in the diminished absorption of nutrients, increased intestinal permeability, and sepsis in patients with thermal injury.

Sign in / Sign up

Export Citation Format

Share Document