A-253

The A-253 cell line is a human epithelial cell line derived from an epidermoid carcinoma of the submaxillary salivary gland. This cell model is widely used to study salivary gland pathophysiology, especially in the context of autoimmune conditions like Sjögren’s syndrome (SS). SS is a chronic autoimmune disease affecting exocrine glands, leading to symptoms of dry mouth (xerostomia) and dry eyes (keratoconjunctivitis sicca), as well as glandular tissue damage. In A-253 cells, studies have shown that certain autoantibodies present in SS patients, such as anti-Ro and anti-La, can trigger apoptosis through both intrinsic and extrinsic pathways. This occurs via interactions with Fc gamma (Fcγ) receptors on the cell surface, leading to internalization of these autoantibodies and activation of apoptotic signaling cascades involving caspases, specifically caspase-3, caspase-8, and caspase-9.

Research has also explored the effects of herbal polysaccharides on the A-253 cell line, particularly from Dendrobium officinale, which has shown potential protective effects against apoptosis induced by tumor necrosis factor-alpha (TNF-α). When pre-treated with these polysaccharides, A-253 cells exhibit a reduction in TNF-α-induced apoptotic markers, such as decreased reactive oxygen species (ROS) production, preserved mitochondrial membrane potential, and reduced translocation of nuclear factor-kappa B (NF-κB) to the nucleus. These observations suggest a potential role for such polysaccharides in mitigating immune-related damage in salivary glands and in providing a novel therapeutic angle for autoimmune diseases like SS, where excessive apoptosis contributes to glandular dysfunction and inflammation.

Overall, A-253 cells serve as an important model for understanding salivary gland cell apoptosis and the molecular mechanisms underlying autoimmune salivary gland damage. This includes the study of caspase activation, mitochondrial destabilization, and pro-inflammatory cytokine effects, which are relevant both in Sjögren’s syndrome and in broader investigations into epithelial cell resilience and immune interactions.

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.