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UK Health Security Agency

Cell Line Profile PNT2

(ECACC catalogue no. 95012613)

Purchase Cell line

 

Cell line history

The human cell line PNT2 originated from the normal prostate tissue of a deceased 33 year old male. A plasmid containing a Simian virus genome with a defective replication origin (SV40 ori-) was used to immortalise the cell line. The Simian virus is commonly used to immortalise mammalian epithelial cells (Cussenot et al, 1991). In the case of PNT1A and PNT2, the large T protein was used as the immortalising agent and liposomes were used as vectors (Cussenot et al, 1991). The use of SV40-immortalised cell lines instead of hTERT-immortalised cell lines is regarded to be more appropriate for research into carcinogenesis (Toouli et al, 2002).

  

48 hours post resuscitation                 Prior to cryopreservation

Key characteristics

PNT2 cells are attached cell lines which grow with an epithelial morphology. PNT2 expresses cytokeratin (CK) 8, 18, and 19, members of the keratin family. The keratin family are responsible for maintaining the structure of epithelial cells. CK8 and CK18 are assembled together as partners, and function as plasminogen receptors (Gonias et al, 2001). CK19 is expressed in the periderm (RefSeq, 2008), and is a feature of differentiated luminal cells.

Applications

PNT2 was established by immortalisation of normal adult prostatic epithelial cells by transfection with a plasmid containing SV40 genome with a defective replication origin. These cells are widely used in cancer research as a non-malignant prostate model. They express cytokeratin (CK) 8, 18, and 19, members of the keratin family, which are responsible for maintaining the structure of epithelial cells. They have supported studies distinguishing cancerous vs. normal cells, measuring cellular mechanics via Atomic Force Microscopy (AFM), and investigating how sodium channel expression can enhance prostate cancer invasiveness.

Culture tips

The media that PNT2 should be cultured in is RPMI 1640 + 2mM Glutamine + 6-10% Foetal Bovine Serum (FBS) at 37°C + 5% CO2. The cells should be seeded at around 2-4 x 104 cells/cm2 and should be passaged when the subculture is between 70-80% confluent using 0.25% trypsin or trypsin/EDTA. These cells should be handled under laboratory containment level 2.

Related cell lines

ECACC catalogue number

Description

PNT2 (Serum free)

07042701

A derivative of PNT2 adapted to grow in serum free media.

PNT1A

95012614

35 year old male human prostate epithelial cells immortalised by SV40 ori-, presenting the phenotype of differentiated luminal prostatic cells with the expression of cytokeratin 8 and 18, and vimentin. Cytokeratin 14 is not expressed. These cells are non-tumourigenic in mice

PNT1A (Serum free)

07052901

A derivative of PNT1A adapted to grow in serum free media.

Key references

  1. Cussenot, O., et al., Immortalization of Human Adult Normal Prostatic Epithelial Cells by Liposomes Containing Large T-SV40 Gene. The Journal of Urology, 1991. 146(3): p. 881-886.

  2. Toouli, C.D., et al., Comparison of human mammary epithelial cells immortalized by simian virus 40 T-Antigen or by the telomerase catalytic subunit. Oncogene, 2002. 21: p. 128.

  3. Gonias, S.L., T.A. Hembrough, and M. Sankovic, Cytokeratin 8 functions as a major plasminogen receptor in select epithelial and carcinoma cells. Front Biosci, 2001. 6: p. D1403-11.

  4. RefSeq. KRT19 Keratin 19 (human). 2008 8 Apr 2018; Available from: https://www.ncbi.nlm.nih.gov/gene/3880.

  5. Baker, M.J., et al., Discrimination of prostate cancer cells and non-malignant cells using secondary ion mass spectrometry. Analyst, 2008. 133(2): p. 175-179.

  6. Faria, E.C., et al., Measurement of elastic properties of prostate cancer cells using AFM. Analyst, 2008. 133(11): p. 1498-1500.

  7. Smith, P., et al., Sodium channel protein expression enhances the invasiveness of rat and human prostate cancer cells. FEBS Letters, 1998. 423(1): p. 19-24.