General Cell Collection ECACC No. 05092804 Cell Line Name VP229 Keywords Human Breast Cancer 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. Species Human Tissue Breast Morphology Epithelial Sub Culture Routine Split sub-confluent cultures (70-80%) 1:2 i.e. seeding at 2-5x10,000 cells/cm� 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. 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. Karyotype Mean no. 74, modal no. 66, range 55-134 Depositor Dr Suet-Feung Chin, Cancer Genomics Program, Hutchnson/MRC Research Centre, Cambridge, UK Originator No Country UK References McCallum & Lowther, 1996. Long-term culture of primary breast cancer indefined medium. Breast Cancer Research and Treatment 39:247-259. Additional Literature Report Not Available Additional Bibliography Not Available Research Council Deposit No Release Conditions No DNA Available from Stock No The ECACC collections represent deposits of cell cultures from world-wide sources. While every effort is made to ensure details distributed by ECACC are accurate, ECACC 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 ECACC does not guarantee the passage number stated will be the passage number received by the customer. Cell lines supplied by ECACC 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 ECACC Terms & Conditions of Supply for more information.   Delivery State Price Code - C Frozen -  �295.00   Growing -  �365.00   DNA -  See Price List and manual Order Form.     Quantity Required    Prices & Ordering Click the 'UPDATE' button to refresh the price for the quantity of packs required. Click the Basket button to add this product to your order.    Net Total           �295.00