Human Cell Lines-Cell Lines-InCellGene LLC.-CellLines,Biomedicine reasearch Product,Cell Biology,Cell Culture,293T Cell,Media

Permits and Restrictions	View Permits View Restrictions
Organism	Homo sapiens, human
Tissue	mammary gland/breast; derived from metastatic site: pleural effusion
Product Format	frozen
Morphology	epithelial
Culture Properties	adherent
Biosafety Level	1 Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Disease	adenocarcinoma
Age	51 years
Gender	female
Ethnicity	Black
Applications	This cell line is a suitable transfection host.
Storage Conditions	liquid nitrogen vapor phase

Karyotype	modal number = 64; range = 60 to 67. The cell line is aneuploid human, presumably female (X, abnormal X) with most chromosome counts in the hypotriploid range.; Normal chromosomes X, N2, N3, N7, N8, N10, and N22 are clearly under-represented due to their involvement in the formation of the many marker (19) chromosomes present in this cell line.; A normal chromosome N1 (or two) is identified in each karyotype, but, in addition, regions of chromosome N1 are also present in five different marker chromosomes.; Variation is evident in the normal and marker chromosome copy number from karyotype to karyotype.

Derivation	The MDA-MB-468 cell line was isolated in 1977 by R. Cailleau, et al., from a pleural effusion of a 51-year-old Black female patient with metastatic adenocarcinoma of the breast.
Clinical Data	51 years Although the tissue donor was heterozygous for the G6PD alleles, the cell line consistently showed only the G6PD A phenotype. Black female
Antigen Expression	Blood Type AB; HLA Aw23, Aw30, B27, Bw35, Cw2, Cw4 (patient)
Receptor Expression	epidermal growth factor (EGF); transforming growth factor alpha (TGF alpha)
Tumorigenic	Yes
Effects	Yes, in nude mice inoculated subcutaneously with 10(7) cells. (Tumors developed within 21 days at 100% frequency (5/5).)
Comments	Although the tissue donor was heterozygous for the G6PD alleles, the cell line consistently showed only the G6PD A phenotype. There is a G -> A mutation in codon 273 of the p53 gene resulting in an Arg -> His substitution. EGF receptor is present at 1 X 106 per cell.

Complete Growth Medium	The base medium for this cell line is ATCC-formulated Leibovitz''''''''s L-15 Medium, Catalog No. 30-2008. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%. (Note: The L-15 medium formulation was devised for use in a free gas exchange with atmospheric air. A CO2 and air mixture is detrimental to cells when using this medium for cultivation)
Subculturing	Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product. Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C. Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended Medium Renewal: 2 to 3 times per week
Cryopreservation	Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Culture Conditions	Atmosphere: air, 100% Temperature: 37°C