skip to main content
UK Health Security Agency

VP229

VP229

Catalogue No.

05092804

Cell Line Name

VP229

Cell Line Description

Derived from a breast cancer biopsy removed from a 47 year-old female. The histology of the breast cancer biopsy was defined as Ductal Grade III, tumour cells did not stain positive for oestrogen receptors. The VP229 cell line was derived from the same individual as the cell line VP267. VP229 was derived from the primary tumour and VP267 was derived 14 months later from a local recurrence of the tumour. The individual was recorded to live for 2 years after surgery performed to remove the primary tumour. This cell line grows slowly and is relatively difficult to culture; please read the sub-culture routine information closely.

Characteristics

Tissue of Origin

Breast

Karyotype

Mean no. 74, modal no. 66, range 55-134

Disease

Carcinoma

Culture Conditions

Cell Type

Epithelial

Subculture Routine

Split sub-confluent cultures (70-80%) 1:2 i.e. seeding at 2-5x10,000 cells/cm² using 0.05% trypsin; 5% CO₂; 37°C. The VP series of breast cancer cell lines are very slow growing and relatively difficult to grow. Fresh EGF (final concentration 10ng/ml) should be added to the medium at each subculture. The subculture split ratio should be kept low at 1:2. Remove the cells with trypsin, it is important to inactivate the trypsin with an equal volume of trypsin inhibitor (the culture medium contains too low a serum content to inactivate the trypsin). To remove all traces of trypsin and trypsin inhibitor pellet the cells by centrifugation and resuspend the cells in a small volume of fesh medium (5ml for a 25cm² flask, 15ml for a 75cm² flask and 25ml for a 175cm² flask). Seed the cells into a new flask in the low volume used to resuspend the cells, leave untouched until the cells have adhered to the flask. Once cells have adhered add more medium to the flask to top up the volume. Change the medium 1-2 times a week until they reach 70-80% confluency. Upon resuscitation of the cell line remove the cryoprotectant (DMSO) by pelleting the resuscitated cells by centrifugation and resuspending in fresh medium. This cell line is relatively difficult to culture, previous experience of the culture of breast epithelial cells in reduced serum conditions is recommended.

Culture Medium

MEBM (Clonetics, CC-3151) + MEGM SingleQuots (Clonetics, CC-4136) + 2mM Glutamine + 2% Foetal Bovine Serum (FBS). Fresh EGF (final concentration of 10ng/ml) should be added to the medium at each subculture.

Growth Mode

Adherent

Additional Info

Depositor

Dr Suet-Feung Chin, Cancer Genomics Program, Hutchnson/MRC Research Centre, Cambridge, UK

Country of Origin

United Kingdom

Hazard Group (ACDP)

Hazard Group (ACDP) 2

Applications

References

McCallum & Lowther, 1996. Long-term culture of primary breast cancer indefined medium. Breast Cancer Research and Treatment 39:247-259.

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: VP229 (ECACC 05092804).

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.