CaCo-2

Caco-2 cells serve as an advanced in vitro model for the human intestinal barrier, primarily due to their differentiation into a cell monolayer that closely resembles the enterocytes lining the small intestine. When culturing the Caco2 cell line on tissue culture filter inserts with polycarbonate filters, Caco-2 cells undergo spontaneous differentiation. The differentiation of Caco2 cells results in the expression of specialized cell types, complete with microvilli, enzymes, and transporters, paralleling the complex features and mechanisms found in an in vivo situation.

In the context of intestinal absorption studies models, Caco-2 cells, which were derived from a human colorectal adenocarcinoma patient, are instrumental due to their ability to develop high TEER values, signifying intact tight junctions and epithelial barrier function. These properties are crucial for assays like the cholesterol efflux assay and investigations into cellular transport, including the movement of lipid nanoparticles and the detection of protein interactions.

Caco-2 cells are pivotal for intestinal absorption studies, providing a reliable in vitro approximation of the intestinal epithelium. Mimicking intestinal enterocytes, these cells facilitate analyses of oral drug absorption by simulating the intestinal barrier. Researchers utilize Caco-2 cells to predict how substances traverse the intestinal mucosa, which is essential for the pharmacokinetic profiling of oral medications. Furthermore, they are a key tool in investigating intestinal cholesterol uptake, homeostasis and transport, which are vital processes for understanding lipid metabolism and associated diseases.

Caco-2 cells remain a cornerstone in colon carcinoma and toxicology research, not only for their relevance to human gastrointestinal studies but also for their role in providing detailed insights into the biliary pathway, the metabolism of xenobiotics within the colon, cancer and toxicology research.

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.