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Intestinal Epithelium

The Intestinal Epithelium is the thin, protective layer of cells that lines the interior of the small and large intestines.
It plays a crucial role in nutrient absorption, immune function, and maintaining the gut barrier.
This epithelium is composed of different cell types, including absorptive enterocytes, secretory goblet cells, and specialized enteroendocrine cells.
Optimizing research on the Intestainl Epithelium is essential for understanding digestive health, intestinal diseases, and developing targeted therapies.
PubCompare.ai's AI-powered platform can help locate the best protocols, compare findings accurately, and enhance reproducibility for your Intestinal Epithelium studies.

Most cited protocols related to «Intestinal Epithelium»

Detailed Isolation Protocols for Diverse Cell Types

For MEF isolation, chimeric embryos were isolated at E13.5, and the head and internal organs were removed. The remaining tissues were physically dissociated and incubated in trypsin at 37° C for 20 min after which cells were resuspended in MEF medium. 24 hours later puromycin (2μg/ml) was added and the cells were expanded for two passages before freezing or plating in doxycline containing media (2μg/ml) for reprogramming experiments. Somatic organs were isolated from 4–6 week-old transgenic mice. Epidermal keratinocytes and intestinal epithelium were isolated and cultured as previously described 6 (link). For mesenchymal stem cells (MSCs) and pro-B cells whole marrow was isolated from the femur and tibia after removal of the condyles at the growth plate by flushing with a syringe and 30-guague needle containing DMEM+5% FBS (Hyclone, Thermo Fisher Scientific). CD19+ pro-B cells were isolated by MACS cell separation (Miltenyibiotec Cat# 130-052-201) following manufactures instructions. Purified B cell subsets were resuspended in IMDM with 15% FCS as well as IL-4, IL-7, SCF (10 ng/ml each, Peprotech), doxycyline (2 μg/ml) and plated on OP9 bone marrow stromal cells (ATCC). Three days later the medium was changed to ESC medium plus Dox. Macrophage cells (CD11b+) from freshly isolated spleen were isolated by MACS cell separation and plated by a similar protocol described for CD19+ pro-B cells. Mesenchymal stem cells were selected through differential plating on tissue culture plates (10cm) for 72h in α-MEM supplemented with 15% FBS. Once plates reached a full monolayer cells were split into 6-well dishes and cultured in the presence of Dox. For isolation of liver cells mice were first perfused with 50 ml HBSS buffer (w/o Ca2+ and Mg2+) then 50 ml HBSS (w/o Ca2+ and Mg2+) containing collagenase (type IV) (Sigma Cat# C5138) (100U/ml). Liver was dissected away from surrounding tissues and dissociated in 10ml DAG media (phenol-red free EMEM Gibco-11054-020 and Bovine serum albumin (BSA) 1g/0.5L) and filtered two times through a sterile 100μM cell strainer. Liver cell preparations were centrifuged at 30 g for 3 minutes at 4 °C and the cells were washed two times with DAG media and then plated on γ-irradiated MEFs in ES media + Dox.

Carey B.W., Markoulaki S., Beard C., Hanna J, & Jaenisch R. (2009). A single-gene transgenic mouse strain for reprogramming adult somatic cells. Nature methods, 7(1), 56-59.

Publication 2009

Bone Marrow Stromal Cells Buffers Cells Cell Separation Chimera Collagenase Condyle Diploid Cell Embryo Epidermis Epiphyseal Cartilage Femur Head Hemoglobin, Sickle Hepatocyte Hyperostosis, Diffuse Idiopathic Skeletal Intestinal Epithelium isolation ITGAM protein, human Keratinocyte Liver Macrophage Marrow Mesenchymal Stem Cells Mice, Transgenic Mus Needles Pro-B Lymphocytes Puromycin Serum Albumin, Bovine Spleen Sterility, Reproductive Syringes Tibia Tissues Trypsin

Screening Bioactive Molecules for Drug-Likeness

In ADME processes, OB is one of the most important pharmacokinetic parameters (18 (link)). High OB is often a key indicator to determine the DL of bioactive molecules. For TCM formulations, the failure of most of the ingredients to reach the protein target sites of particular cells is due to a lack of appropriate pharmacologic properties, especially OB. Molecules with OB ≥30% were considered to have good OB in the present study.
In the early stages of drug development, DL evaluation helps to screen out excellent compounds (19 (link)) and increases the ‘hit rate’ of drug candidates. Therefore, the DL of molecules in YCHD was assessed using the Tanimoto coefficient in the present study (20 (link)) using the following formula:
Where × is the molecular descriptor of YCHD based on Dragon software (http:www.talete.mi.it/products/dragon_description.htm) and y is the average descriptor of all drugs in the Drugbank database. The average DL Index of all drugs in the Drugbank database is 0.18, which indicates a high DL. Thus in our study, active molecules were defined as those with a DL Index ≥0.18.
The intestinal epithelial permeability can be investigated using Caco-2 cells (21 (link)). Orally administered drugs are absorbed mainly through intestinal epithelial cells. Therefore, simulation of drug transport across the monolayers of small-intestinal epithelial cells is crucial for the prediction of drug absorption. The permeability of epithelial cells of ingredients in Chinese herbal medicines was predicted using the TCMSP database. It was considered that molecules with Caco-2 >-0.40 had good permeability in the small-intestinal epithelium.
Hence, the selected candidate molecules had to meet the requirements of OB ≥30%, DL ≥0.18 and Caco-2 >-0.40 for further analyses.

Huang J., Cheung F., Tan H.Y., Hong M., Wang N., Yang J., Feng Y, & Zheng Q. (2017). Identification of the active compounds and significant pathways of yinchenhao decoction based on network pharmacology. Molecular Medicine Reports, 16(4), 4583-4592.

Publication 2017

Caco-2 Cells Chinese Epithelial Cells Intestinal Epithelium Intestines Medicines, Herbal Permeability Pharmaceutical Preparations Protein Targeting, Cellular

Triculture Model of Intestinal Epithelium

A number of in vitro models have been used to study the toxicity and biokinetics of pharmaceuticals and chemicals in the GIT. The most commonly used model employs Caco-2 cells (immortal human colonic epithelial) cells, which after culture for 2–3 weeks differentiate into cells with markers and morphological characteristics of small intestinal epithelial enterocytes [64 –66 (link)]. While this may be a reasonable choice for many situations, the epithelium of the small intestine is more complex, and in order to more accurately emulate this structure, a variety of modifications have been added. The intestinal mucosa is normally protected by a layer of mucus produced by both goblet cells and submucosal glands (Brunner’s glands, limited mostly to the duodenum) [67 (link)]. It is therefore appropriate to modify the in vitro model to include mucus secreting cells. To this end, HT29-MTX cells, an immortal human cell line that resembles intestinal goblet cells and secretes mucus, is often co-cultured with Caco-2 cells [66 (link)–69 (link)]. Finally, in the Peyer’s patches and other lymphoid-associated epithelium of the small intestine, specialized cells called Microfold- or M-cells are present. These cells engulf and translocate samples of the contents of the intestinal lumen to lymphocytes in the submucosa below, thereby providing continuous antigenic surveillance of the intestinal contents [33 (link)]. It has also recently been shown that M-cells can play an important role in translocation of iENMs in in vitro intestinal epithelial models [33 (link)]. It has previously been shown that differentiated Caco-2 cells can be induced by factors released from another cell line, Raji B (a human B lymphocyte) to differentiate into cells resembling M-cells [70 (link), 71 (link)]. Thus, when Raji B cells are added to the basolateral compartment of a transwell system in which matured caco-2 cells reside on the transwell membrane above, some of the Caco-2 cells are induced to differentiate into M-like cells. The complete hybrid triculture model utilized in our methodology, illustrated in Fig. 5a, has previously been described and characterized and includes cells with morphology and markers consistent with the three primary cells of the intestinal epithelium: enterocytes, goblet cells and M-cells [37 (link)–41 (link)]. Because it represents a reasonably realistic hybrid model of the complete intestinal epithelium, this model was adopted for the proposed integrated methodology. Specifically, we employed the protocol reported by Mahler et al. [37 (link)] for development of our triculture system. Such a physiologically relevant model is well suited to the study of biokinetics and intestinal toxicity of iENMs. However other similar advanced models could also be used.
Details of the methods employed for development, characterization and validation of the triculture model, including protocols for creating the system, measurement of transepithelial electrical resistance (TEER), immunofluorescence staining and imaging for morphological characterization and TEM characterization are provided in Additional file 1.

DeLoid G.M., Wang Y., Kapronezai K., Lorente L.R., Zhang R., Pyrgiotakis G., Konduru N.V., Ericsson M., White J.C., De La Torre-Roche R., Xiao H., McClements D.J, & Demokritou P. (2017). An integrated methodology for assessing the impact of food matrix and gastrointestinal effects on the biokinetics and cellular toxicity of ingested engineered nanomaterials. Particle and Fibre Toxicology, 14, 40.

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Publication 2017

Antigens B-Lymphocytes Brunner Glands Caco-2 Cells Cell Lines Cells Colon Cultured Cells Duodenum Enterocytes Epithelial Cells Epithelium Goblet Cells Homo sapiens HT29 Cells Hybrids Immunofluorescence Intestinal Contents Intestinal Epithelium Intestinal Mucosa Intestines Intestines, Small Lymph Lymphocyte M Cells Mucus Peyer Patches Pharmaceutical Preparations Resistance, Electrical Tissue, Membrane Translocation, Chromosomal

Measuring ROS Levels in Drosophila Intestine

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Publication 2010

Cells Diptera Genotype Hypersensitivity Intestinal Epithelium Intestines Microscopy, Confocal Tissues

Culturing Neonatal Swine Intestinal Epithelial Cells

PIE cells are intestinal non-transformed cultured cells originally derived from intestinal epithelia isolated from an unsuckled neonatal swine (16 (link)). When PIE cells are cultured, they assume a monolayer with a cobblestone and epithelial-like morphology and with close contact between cells (14 (link), 16 (link), 17 (link)). PIE cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen Corporation, Carlsbad, CA, USA) supplemented with 10% fetal calf serum, 100 U/ml streptomycin, and 100 mg/ml penicillin at 37°C in an atmosphere of 5% CO₂. PIE cells grow rapidly and are well adapted to culture conditions even without transformation or immortalization (17 (link)–19 (link)).

Albarracin L., Kobayashi H., Iida H., Sato N., Nochi T., Aso H., Salva S., Alvarez S., Kitazawa H, & Villena J. (2017). Transcriptomic Analysis of the Innate Antiviral Immune Response in Porcine Intestinal Epithelial Cells: Influence of Immunobiotic Lactobacilli. Frontiers in Immunology, 8, 57.

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Publication 2017

Atmosphere Cells Cultured Cells Eagle Fetal Bovine Serum Infant, Newborn Intestinal Epithelium Intestines Penicillins Pigs Streptomycin

Most recents protocols related to «Intestinal Epithelium»

Expression Analysis of Chl1 in Intestinal Cell Populations

Not available on PMC !

Example 2

Next, the expression of Chl1 was confirmed using various tissues or cells.

(B) The intestinal epithelium (EpCAM-positive CD45-negative), fibroblasts (COL1a2-GFP-positive CD45-negative podoplanin-positive), macrophages (F480-positive CD11b-positive), CD4-positive T cells, B cells (CD19-positive B220-positive), and lamina propria cells of the large intestine (whole colon cells) were isolated in the same way as above. RNA was purified from each cell using TRIZOL (Thermo Fisher Scientific Inc./Invitrogen: 15596018) and subsequently reverse-transcribed using VILO (Thermo Fisher Scientific Inc./Invitrogen: 11755500). The expression analysis of Chl1 was conducted using Universal Probe Library (Roche Life Science) and LightCycler™ 480 system (Roche Life Science). Comparison with the expression of Gapdh is shown (n=3). The results were as shown in FIG. 2.

As shown in FIG. 2, Chl1 exhibited high expression in fibroblasts, particularly, fibroblasts at the acute phase (“Acute”) and chronic phase “Chronic” of induced inflammation. This suggested that Chl1 may be preferably used as a marker for acute and chronic inflammation.

US11866497B2. Antibody to fibrosis-related molecule and medical application thereof (2024-01-09). The University of Tokyo [JP]. Inventors: Yosuke Kurashima [JP], Hiroshi Kiyono [JP].

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Patent 2024

B-Lymphocytes CD4 Positive T Lymphocytes cDNA Library Cells COL1A2 protein, human Colon Fibroblasts GAPDH protein, human Inflammation Intestinal Epithelium ITGAM protein, human Lamina Propria Large Intestine Macrophage TACSTD1 protein, human Tissues trizol

Histological Scoring of Colon Inflammation

4 µm paraffin sections from the fixed colon (caecum and proximal part) were cut serially, mounted onto glass slides, and deparaffinized. The colon sections were stained with hematoxylin and eosin by the Core Facility (IZKF) of the RWTH Aachen University. Blinded histological scoring was performed using a standard microscope, based on TJL method as described previously²⁶. Each colon section was scored for the four general criteria: severity, degree of hyperplasia, degree of ulceration, if present, and percentage of area involved. A subjective range of 1–3 (1 = mild, 2 = moderate, 3 = severe) was used for the first three categories. Severity: Focally small or widely separated multifocal areas of inflammation limited to the lamina propria were graded as mild lesions (1). Multifocal or locally extensive areas of inflammation extending to the submucosa were graded as moderate lesions (2). If the inflammation extended to all layers of the intestinal wall or the entire intestinal epithelium was destroyed, lesions were graded as severe (3). Hyperplasia: Mild hyperplasia consisted of morphologically normal epithelial lining that was at least twice as thick (length of crypts) as adjacent or control mucosa. Moderate hyperplasia was characterized by the epithelial lining being two- or three-times normal thickness, cells were hyperchromatic, numbers of goblet cells were decreased, and scattered individual crypts developed an arborizing pattern. Severe hyperplastic regions exhibited markedly thickened epithelium (four or more times normal thickness), marked hyperchromasia of cells, few to no goblet cells, a high mitotic index of cells within the crypts, and numerous crypts with arborizing pattern. Ulceration was graded as: 0 = no ulcer, 1 = 1–2 ulcers (involving up to a total of 20 crypts), 2 = 1–4 ulcers (involving a total of 20–40 crypts), and 3 = any ulcers exceeding the former in size. A 10% scale was used to estimate the area involved in the inflammatory process. 0 = 0%, 1 = 10–30%, 2 = 40–70%, 3 = > 70%.

Subramanyam S.H., Hriczko J.T., Pappas A., Schippers A., Wagner N., Ohl K, & Tenbrock K. (2023). Tofacitinib fails to prevent T cell transfer colitis in mice but ameliorates disease activity. Scientific Reports, 13, 3762.

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Publication 2023

Cecum Cells CFC1 protein, human Colon Eosin Epithelium Goblet Cells Hyperplasia Inflammation Intestinal Epithelium Intestines Lamina Propria Microscopy Mucous Membrane Paraffin Ulcer

Quantifying Intestinal Cell Proliferation

7 dpf larvae were immersed overnight in 100 μg/mL 5-ethynyl-2′-deoxyuridine (EdU) solution (A10044; Invitrogen) for 16 h before termination of the experiment at 8 dpf. Larvae were fixed in 4% paraformaldehyde for 24 hr at 4°C, processed for paraffin embedding, and cut into 7-μm sections. For EdU detection, slides were processed according to the Click-iT EdU Cell Proliferation Assay Kit (C35002; Molecular Probes). Samples were imaged on a Nikon Eclipse TE 2000-V inverted microscope equipped with a Photometrics Coolsnap camera. EdU-labeled nuclei within the intestinal epithelium were counted over 30 serial 7-μm sections beginning at the esophageal-intestinal junction and proceeding caudally into the bulb. Analysis of this extended region was necessary because of the stochastic patterns of cell proliferation. The absolute numbers of labeled cells varied between trials. Despite these differences in the absolute numbers of labeled cells, the proportional trends of proliferating cells between treatments were consistent and reproducible between trials.

Banse A.V., VanBeuge S., Smith T.J., Logan S.L, & Guillemin K. (2023). Secreted Aeromonas GlcNAc binding protein GbpA stimulates epithelial cell proliferation in the zebrafish intestine. Gut Microbes, 15(1), 2183686.

Publication 2023

Biological Assay Cell Nucleus Cell Proliferation Deoxyuridine Intestinal Epithelium Intestines Larva Medulla Oblongata Microscopy Molecular Probes paraform Somatostatin-Secreting Cells

Quantifying Intestinal MUC2 and Bacterial Encroachment

Three images from 4 mice per group were used to quantify MUC2 intensity and bacterial encroachment. For MUC2 intensity, ImageJ (NIH) was used to set a threshold and mask for each image, and pixel intensity was measured using ImageJ measuring tool. Bacterial encroachment was measured as the distance between the closets bacteria to the intestinal epithelium using the ImageJ measuring tool.

Hill A.A., Kim M., Zegarra-Ruiz D.F., Chang L.C., Norwood K., Assié A., Wu W.J., Renfroe M.C., Song H.W., Major A.M., Samuel B.S., Hyser J.M., Longman R.S, & Diehl G.E. (2023). Acute high-fat diet impairs macrophage-supported intestinal damage resolution. JCI Insight, 8(3), e164489.

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Publication 2023

Bacteria Intestinal Epithelium Mice, House MUC2 protein, human

Conditioned Medium from L. rhamnosus Adhesion

Since L. rhamnosus is not able to cross the intestinal epithelium in physiological conditions, its anti-inflammatory activity was tested using the conditioned medium following adhesion to intestinal epithelium. L. rhamnosus adhesion to the intestinal epithelium in vitro model was performed as described by Candela and colleagues [17 (link)]. Before adhesion assay, the intestinal epithelium in vitro model was thoroughly washed with Hank’s Balanced Salt Saline (HBSS) and incubated for 1 h at 37 °C with cell culture medium without antibiotics. Subsequently, 8.6 × 10⁸ CFU/mL (equivalent to a Multiplicity of Infection (MOI) of about 200 bacteria per Caco-2 cell) was added to the apical compartment in cell culture medium without antibiotics and incubated for 3 h in controlled atmosphere incubator (37 °C, 85% relative humidity and 5% CO₂). Then, Caco-2 monolayers were extensively washed with HBSS to eliminate non-adhered bacteria. Adhered bacteria were left to grow for 24 h and the resulting conditioned medium (i.e., media containing components and metabolic products secreted by L. rhamnosus) was sterile-filtered and stored at −80 °C for further application.

Murzilli S., Mirone V., Micheletto M., Tedesco E., Maira G.D., Benetti F, & Vanelli A. (2023). Evaluation of the Immunomodulatory Effects of a Probiotics and Natural Extract-Based Formulation in Bacterial-Induced Prostatitis. Life, 13(2), 389.

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Publication 2023

Anti-Inflammatory Agents Antibiotics Atmosphere Bacteria Bacterial Infections Biological Assay Caco-2 Cells Cell Culture Techniques Cells Culture Media Culture Media, Conditioned Epithelium Humidity Intestinal Diseases Intestinal Epithelium Iodine physiology Saline Solution Salts Strains

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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.

Dulbecco modified eagle medium (dmem) by Thermo Fisher Scientific

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DMEM (Dulbecco's Modified Eagle's Medium) is a cell culture medium formulated to support the growth and maintenance of a variety of cell types, including mammalian cells. It provides essential nutrients, amino acids, vitamins, and other components necessary for cell proliferation and survival in an in vitro environment.

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Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.

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What are the main cell types found in the Intestinal Epithelium?

The Intestinal Epithelium is composed of several key cell types, including: 1. Absorptive enterocytes: These cells are responsible for nutrient absorption from the intestinal lumen. 2. Secretory goblet cells: These cells produce mucus, which helps lubricate and protect the intestinal surface. 3. Enteroendocrine cells: These specialized cells secrete hormones that regulate various digestive functions. 4. Paneth cells: Found in the small intestine, these cells secrete antimicrobial substances to help maintain the gut barrier. Understanding the different cell types and their functions within the Intestinal Epithelium is crucial for researching digestive health and developing targeted therapies.

What are some common challenges in studying the Intestinal Epithelium?

Some of the key challenges in studying the Intestinal Epithelium include: 1. Maintaining the delicate structure and function of the epithelium in vitro. 2. Accurately modeling the complex interactions between the epithelium, gut microbiome, and immune system. 3. Isolating and characterizing the different cell types within the epithelium. 4. Reproducibly measuring epithelial barrier function and nutrient absorption capabilities. PubCompare.ai can help address these challenges by providing a comprehensive database of protocols and studies related to the Intestinal Epithelium. The platform's AI-powered analysis can identify the most effective methods and highlight critical insights to enhance the accuracy and reproducibility of your research.

How can PubCompare.ai assist in Intestinal Epithelium research?

PubCompare.ai's AI-powered platform can be a valuable tool for Intestinal Epithelium research in several ways: 1. Efficienctly screen protocol literature: The platform can help you quickly identify the most relevant and effective protocols for your specific research goals related to the Intestinal Epithelium. 2. Leverage AI to pinpoint Critical Insights: PubCompare.ai's AI-driven analysis can highlight key differences in protocol effectiveness, enabling you to choose the best option for reproducibility and accuracy. 3. Identify the most effective protocols: The platform can help researchers identify the optimal methods and products for Intestinal Epithelium studies, improving the reliability and comparability of your findings. By leveraging PubCompare.ai's capabilities, you can enhance the efficiency, accuracy, and reproducibility of your Intestinal Epithelium research, leading to more impactful discoveries and advancements in digestive health and targeted therapies.

More about "Intestinal Epithelium"

The intestinal epithelium is a crucial component of the digestive system, serving as a protective barrier and playing a vital role in nutrient absorption, immune function, and gut health.
This thin layer of cells lining the small and large intestines is composed of various cell types, including absorptive enterocytes, secretory goblet cells, and specialized enteroendocrine cells.
Optimizing research on the intestinal epithelium is essential for understanding digestive disorders, developing targeted therapies, and promoting overall gastrointestinal wellbeing.
Researchers can leverage cutting-edge tools and techniques to study this complex tissue, such as utilizing Transwell inserts for in vitro modeling, employing DNase I to isolate nucleic acids, and employing the BCA protein assay kit for quantifying protein levels.
Additionally, the use of cell culture media like DMEM, supplemented with FBS and antibiotics like penicillin/streptomycin, can provide a conducive environment for intestinal epithelial cell growth and experimentation.
The Agilent 2100 Bioanalyzer can also be leveraged to analyze the quality and quantity of RNA extracted from intestinal epithelial samples.
For studies aimed at understanding the effects of specific compounds on the intestinal epithelium, the use of Tamoxifen, a selective estrogen receptor modulator, may provide insights into the tissue's response to hormonal influences.
Furthermore, the MITO+ Serum Extender can be employed to enhance mitochondrial function and energy production in intestinal epithelial cells, potentially contributing to a better understanding of their metabolic processes.
By integrating these specialized tools and techniques, researchers can optimize their investigations of the intestinal epithelium, leading to advancements in our understanding of digestive health, the development of innovative therapies, and the promotion of overall gastrointestinal wellbeing.
The field of intestinal epithelium research holds immense potential for unlocking new frontiers in the realm of digestive and overall health.