| Field | Specification |
|---|---|
| Species |
The PK(15) cell line, derived from PK-2A, a cell line established in 1955 from the kidney of an adult pig, is infected with the porcine type-C oncovirus (formerly known as porcine endogenous retrovirus, PERV), which is classified as a risk group 2 agent. 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 the PK(15) cell line were described as defective and non-infectious for a variety of mammalian cell lines, including a human cell line, leading to its classification as a risk group 1 cell line. However, subsequent studies demonstrated that human 293 cells could be productively infected by the cell-free supernatant of PK(15) cells. This finding resulted in the reclassification of the PK(15) cell line by the German Central Commission for Biological Safety (ZKBS) in November 2018. PCR analyses revealed that the transmitted viruses belonged to the polytropic subtypes PERV-A and PERV-B. Additionally, 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 as a suitable host for transfection applications. Due to its adherent growth properties, it is highly valuable in various research and experimental settings.
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- Viruses: PCV1 (Porcine circovirus 1) positive, PCV2 negative, PCV3 negative
- Virus susceptibility: 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
- Virus resistance: Poliovirus 2
- Reverse transcriptase: Positive
- cultureMedium: EMEM (MEM Eagle), w: 2 mM L-Glutamine, w: 2.2 g/L NaHCO3, w: EBSS (Cytion article number 820100a)
- supplements: Supplement the medium with 10% FBS and 1% NEAA
- dissociationReagent: Accutase
- subculturing: 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.
- seedingDensity: 2 x 104 cells/cm2
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: Allow the cells to recover from the freezing process for at least 24 to 48 hours.
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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