IEC-18 Cells

Categories: Rat cell lines Description: The IEC-18 cell line is a non-transformed epithelial cell line derived from the crypt cells of rat small intestine. These cells have been shown to effectively model the physiological properties of the small intestinal epithelium, particularly with respect to chloride ion (Cl-) transport. Chloride channels in IEC-18 cells exhibit distinct types of conductances that respond to various stimuli such as cell swelling, increased intracellular calcium (Ca2+), and elevated cyclic AMP (cAMP). For instance, swelling-activated Cl- currents in IEC-18 cells are characterized by outward rectification and voltage independence. Moreover, IEC-18 cells express cystic fibrosis transmembrane conductance regulator (CFTR) channels, evidenced by the presence of cAMP-activated Cl- conductance which can be inhibited by glibenclamide and 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB), but not affected by DIDS. IEC-18 cells have also been used to explore cell survival mechanisms under detachment-induced stress, known as anoikis. Research indicates that prostaglandin E2 (PGE2) can promote cell viability and aggregation in detached IEC-18 cells through cAMP-mediated signaling pathways. This protection from anoikis is associated with the activation of adenylate cyclase and protein kinase A (PKA), enhancing cell adhesion and viability even in suspended states. Such findings are significant for understanding inflammation-related processes and potential contributions to carcinogenesis in intestinal tissues. Furthermore, IEC-18 monolayers have been employed to study the transport of various molecules across the intestinal barrier. Compared to the Caco-2 cell line, IEC-18 cells provide a more accurate model for passive transcellular and paracellular transport due to their structural similarities to small intestine crypt cells. Unlike Caco-2 cells, which possess significant active transport capabilities, IEC-18 cells demonstrate minimal carrier-mediated transport, making them a more suitable choice for analyzing the passive permeability of hydrophilic macromolecules. Organism: Rat Tissue: Small intestine, ileum Disease: Normal Synonyms: IEC 18, IEC18, Intestinal Epithelioid Cell line No. 18 Breed: Charles River Sprague Dawley (CD(SD)) Age: 18-24 days Gender: Unspecified Morphology: Epithelial-like Cell Type: Epithelial cell Growth Properties: Adherent Citation: IEC-18 (Cytion catalog number 305302) Biosafety Level: 1 Ncbi_ Taxid: 10116.0 Cellosaurus Accession: CVCL_0342 Culture Medium: DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a) Supplements: Supplement the medium with 10% FBS Dissociation Reagent: Accutase Subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. Seeding Density: 2 x 104 cells/cm2 Fluid Renewal: 2 times per week Freeze Medium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress. Thawing And Culturing Cells: Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks, for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes. Sterility: Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. Safety Precautions: When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments. Warranty: We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success. Subject To Material Transfer Agreements: If you intend to use Cytion cell lines solely for internal research at a single research site, please complete and sign our Material Transfer Agreement (MTA) and submit it along with your order.For any commercial applications - including but not limited to fee-for-service work, quality control testing, product release, diagnostic use, or regulatory studies - please complete the Intended Use Form so we can prepare a suitable agreement tailored to your project.Please note: The MTA applies only to certain cell lines. If this notice and the MTA document appear on a product page, the agreement is applicable. For cell lines not covered by the MTA, no reference to the agreement will be shown. The MTA is not valid for customers in the Americas, China, or Taiwan. Please contact our U.S. entity to receive the appropriate agreement. Required Product 1: 820300a Required Product 3: 860015.0 Required Product 4: 830100.0