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UK Health Security Agency

DLKP-A5F

DLKP-A5F

Catalogue No.

20082608

Cell Line Name

DLKP-A5F

Cell Line Description

Adriamycin-resistant subpopulation derived from DLKP-A, an Adriamycin-resistant variant of a poorly differentiated lung carcinoma from a 52yr old male. Depositor states the doubling time of about 16 hrs. Parental DLKP displays chromosomal deletions and rearrangements (translocations and isochromosomes). DNA finger-printing analysis of DLKP-A5F confirmed identity as sourced from DLKP-A (data from Depositor).

 

Cell Type - Poorly differentiated squamous, Anchorage-dependent.

Characteristics

Products

DNA fingerprinting from the Depositor confirmed DLKP-A5F to originate from DLKP-A. Subpopulation of DLKP-A isolated by limiting cell dilution and expansion of individual colonies. Expresses PGP. IC50 for Adriamycin 8208 nM, 331-fold more resistant than parent. Cross-resistant to Vincristine (2451-fold compared to parent (DLKP IC50 1.2nM)) and VP16 (64.7-fold compared to control (DLKP IC50 146 nM))

Tissue of Origin

Poorly differentiated lung carcinoma, established from a lymph node biopsy.

Morphology

Epithelial

DNA profile (STR Profile)

Amelogenin: X, X
CSF1PO: 12, 12
D3S1358: 17, 17
D5S818: 10, 11
D7S820: 8, 11
D8S1179: 13, 14
D13S317: 12, 12
D16S539: 10, 12
D18S51: 13, 13
FGA: 20, 20
Penta D: 11, 11
Penta E: 7, 15
TH01: 9.3, 9.3
TPOX: 8, 11
vWA: 17, 17

The Y chromosome could not be detected in this cell line by short tandem repeat (STR)-PCR analysis when tested at ECACC. It is a known phenomenon that due to the increased genetic instability of cancer cell lines the Y chromosome can be rearranged or lost resulting in lack of detection. The cell line is identical to the source provided by the depositor based on the STR-PCR analysis.

Karyotype

Parental DLKP displays chromosomal deletions and rearrangements (translocations and isochromosomes). DNA finger-printing analysis of DLKP-A5F confirmed identity as sourced from DLKP-A.

Applications

Clonal variant of DLKP-A used in drug resistance studies.

Disease

Lung squamous cell carcinoma

Culture Conditions

Cell Type

Poorly differentiated squamous

Subculture Routine

Upon resuscitation count cells and seed at the higher end of the seeding density to establish culture. Spin after thawing to remove DMSO, feed with fresh growth medium the following day. Seed cells between 1-4 x 104 cells/cm2.

 

8% CO2 at 37°C (as medium is a mixture of DMEM/F-12 Ham's). Vented flasks required.

 

Trypsinise and spin in growth medium and feed twice a week with fresh media.

Culture Medium

DMEM:Ham's F-12 (1:1) + 5% Foetal Bovine Serum (FBS)

 

Adriamycin at a final concentration of 2.45 µg/ml can be added 24hrs after the first passage and removed before cryopreservation.

 

Cells are more susceptible to the selecting drug when freshly thawed from liquid nitrogen. It is recommended that the cells are passaged at least three times from thawing before adding selection drug as stated by the Depositor. ECACC will add the selection drug before this time-point as it is unlikely to be passaged three times before cryopreserving.

 

Freeze in 5% DMSO; 45% FBS; 50% growth medium

Growth Mode

Anchorage-dependent

Additional Info

Depositor

Joanne Keenan, Justine Meiller, Martin Clynes, National Institute for Cellular Biotechnology, Dublin City University, Glasnevin, Dublin 9, D09 NR58, Ireland.

Country of Origin

Ireland

Hazard Group (ACDP)

Hazard Group (ACDP) 2

Applications

References

Cleary, I et al. “The multidrug-resistant human lung tumour cell line, DLKP-A10, expresses novel drug accumulation and sequestration systems.” Biochemical pharmacology vol. 53,10 (1997): 1493-502. doi:10.1016/s0006-2952(97)00003-8 PMID: 9260877

Bibliography

Keenan, Joanne et al. “Proteomic analysis of multidrug-resistance mechanisms in adriamycin-resistant variants of DLKP, a squamous lung cancer cell line.” Proteomics vol. 9,6 (2009): 1556-66. doi:10.1002/pmic.200800633 PMID: 19242932

Available Formats

  • Frozen
  • DNA-5µg (100ng/µl)

If use of this culture results in a scientific publication, it should be cited in the publication as: DLKP-A5F (ECACC 20082608).

Unless specified otherwise, at ECACC we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines (Advisory Committee on Dangerous Pathogens) (UK). All cell cultures have the potential to carry as yet unidentified adventitious agents. It is the responsibility of the end user to ensure that their facilities comply with biosafety regulations for their own country.

 

ACDP Guidance: Biological agents: Managing the risks in laboratories and healthcare premises.

The Culture Collections represent deposits of cultures from world-wide sources. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. References where quoted are mainly attributed to the establishment of the cell culture and not for any specific property of the cell line, therefore further references should be obtained regarding cell culture characteristics. Passage numbers where given act only as a guide and Culture Collections does not guarantee the passage number stated will be the passage number received by the customer.

Cultures supplied by Culture Collections are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.