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.

Also Known As:

Species: Human

Tissue: Breast

Growth Properties: Adherent

Morphology: Epithelial

Growth 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.

Subculturing Procedure: Split sub-confluent cultures (70-80%) 1:2 i.e. seeding at 2-5×10,000 cells/cm² using 0.25% 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.

Release Conditions: No

Depositor:

Originator: No

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

Hyperlink to ECACC Cell Line Data Sheet: 05092804