IEC-18

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.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. 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.

3. 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.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. 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.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. 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.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

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.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.