MDCK-SIAT1

The MDCK-SIAT1 cell line is a modified version of the Madin-Darby Canine Kidney (MDCK) cells, engineered to express higher levels of human 2,6-sialyltransferase (SIAT1). This enzyme is responsible for the addition of sialic acid in an alpha-2,6 linkage to galactose on glycoproteins and glycolipids. The modification was performed to increase the expression of alpha-2,6-linked sialic acids, which are the primary receptors for human influenza viruses. This enhancement is critical as it makes the MDCK-SIAT1 cells more similar to the human airway epithelium, which naturally has a high concentration of these receptors. As a result, these cells offer a more physiologically relevant model for studying human influenza viruses and their interactions with potential antiviral compounds.

One of the significant applications of MDCK-SIAT1 cells is in the assessment of influenza virus sensitivity to neuraminidase inhibitors (NAIs), such as oseltamivir. Due to the increased presence of alpha-2,6-linked sialic acids, the MDCK-SIAT1 cells demonstrate improved sensitivity to NAIs compared to unmodified MDCK cells. This makes them an excellent tool for detecting resistance to these inhibitors, especially in low-passage-number clinical isolates of human influenza viruses. The MDCK-SIAT1 cell line allows for more accurate in vitro studies of drug efficacy and viral receptor interactions, providing valuable insights into the development of antiviral therapies and resistance mechanisms.

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.