LLC-PK1

LLC-PK1 cells are a well-established and widely used cell line in biomedical research. These cells were derived from a healthy male pig's kidney, exhibiting typical epithelial morphology. The LLC-PK1 line is polarized and contains tight junctions, making it an ideal model for epithelial tissue.

One of the critical features of LLC-PK1 cells is their ability to produce plasminogen activator, a substance that stimulates fibrinolysis. This property has made LLC-PK1 cells particularly valuable in thrombosis research.

In recent years, plasminogen activator has been included in drugs used in thrombosis therapies since it facilitates the dissolution of small blood clots. In addition to producing plasminogen activators, LLC-PK1 cells produce large amounts of cytokeratin. This characteristic has made them popular for various pharmacologic and metabolic research investigations.

The LLC-PK1 line has been used in drug metabolism, transport, toxicity, and interaction studies. LLC-PK1 cells are also frequently used in permeability assays. The mechanism of uracil transport differs depending on cell lines, with a Na+-independent system on the basolateral membrane in Caco-2 cells and both Na+-dependent and Na+-independent systems on the apical membrane in LLC-PK1 cells.

Compared to other cell lines, LLC-PK1 cells share many characteristics of proximal tubular cells in vivo, including apical membrane microvilli, high activities of apical membrane enzymes, and expression of parathyroid hormone receptors and sodium-dependent glucose transporters. This makes LLC-PK1 cells a valuable tool in renal toxicology studies. Another cell line commonly used in renal toxicology studies is the MDCK cell line. Like LLC-PK1 cells, MDCK cells are epithelial but have characteristics more typical of distal tubular cells.

They express vasopressin, oxytocin, and prostaglandin receptors, which, when stimulated, activate adenylate cyclase. LLC-PK1 and MDCK cell lines proliferate rapidly and can be passaged easily for many generations in monolayer cultures. LLC-PK1 cells are also capable of forming 'domes', fluid-filled blisters resulting from water and solute transport, tight junctions, and adhesion of the cells to the substratum.

In conclusion, the LLC-PK1 cell line is a versatile and valuable tool for biomedical research. It has been widely used in various studies on drug metabolism, drug transport, drug toxicity, drug-drug interactions, renal toxicology, and permeability assays. With its well-established epithelial morphology and plasminogen activator and cytokeratin production, LLC-PK1 cells are an ideal model for epithelial tissue.

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.