PK-15 Cells

The PK(15) cell line, derived from PK-2A, a cell line obtained from the kidney of an adult pig in 1955, is infected with the porcine type-C oncovirus, also known as the porcine endogenous retrovirus (PERV). The host cell genome contains 62 copies of the pol gene, which codes for reverse transcriptase and other proteins.
Initially, the virus particles produced by PK(15) were found to be defective and unable to infect various mammalian cell lines, including a human cell line. However, subsequent studies demonstrated that human 293 cells could be productively infected by the cell-free supernatant from PK(15) cells.
Polymerase chain reaction (PCR) analyses showed that the transmitted viruses belonged to the polytropic subtypes PERV-A and PERV-B.
Furthermore, it was observed that the virus particles produced by the 293 cells were resistant to inactivation by the human complement system. In addition to its virological significance, the PK(15) cell line also serves specific applications as a suitable transfection host.
With its adherent growth properties, the PK(15) cell line proves valuable in various research and experimental settings.

Monolayer, adherent

Male

Epithelial-like

EMEM, w: 2 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: EBSS, w: 1 mM Sodium pyruvate, w: NEAA (Cytion article number 820100c)

Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.

Allow the cells to recover from the freezing process for at least 24 to 48 hours.

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

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.

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.

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

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.

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

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.

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

When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments.

We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success.

A ratio of 1:2 to 1:4 is recommended

Hog cholera, African swine fever, vesicular exanthema of swine, foot and mouth disease (FMDV), vesicular stomatitis (Indiana), vaccinia, reovirus 2, 3, adenovirus 4, 5, coxsackievirus B2, B3, B4, B5, B6

Poliovirus 2