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

Permits and Restrictions	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	43 years
Gender	female
Ethnicity	Caucasian
Applications	This cell line is suitable as a transfection host.
Storage Conditions	liquid nitrogen vapor temperature

Karyotype	This is a hypertriploid human cell line with the modal chromosome number of 84, occurring in 34% of cells. Cells having 80 chromosomes also occurred at a high rate (28%); the higher ploidy cells occurred at 7.3%. This cell line has a very complex chromosome composition. Thirty-five to 40% of chromosomes in a cell complement with a modal chromosome number of 84 consisted of structurally altered marker chromosomes. Several markers are longer than chromosome N1. The origins of most of these markers, however, are not clear. Some markers may have at least three individual chromosome segments. The markers [i.e., ?der(1)t(1;21) (p13;q21) [or ?t(1q21q)], ?del(2) (q13), and t(7pter--cen--?), present in some cells only] were the only ones in which portions of chromosome segments could be identified. Most cells had about three normal X chromosomes and five or more N7. The structurally normal N1, N14 and N17 were generally absent.

Derivation	This cell line was derived by G. Trempe and L. J. Old in 1970 from pleural effusion cells of a patient, a White, Caucasian female, age 43, blood type A+, who had been treated with radiation, steroids, cytoxan and 5-fluorouracil.
Clinical Data	43 years Caucasian female
Antigen Expression	Blood Type A; Rh+; HLA A11, Bw22(+/-), B40, B18
Tumorigenic	Yes
Effects	Yes, in nude mice; forms poorly differentiated adenocarcinoma
Comments	The patient, a White, Caucasian female, age 43, blood type A+, had been treated with radiation, steroids, cytoxan and 5-fluorouracil. No virus particles. Ultrastructural features include microvilli and desmosomes, glycogen granules, large lysosomes, bundles of cytoplasmic fibrils. The SK-BR-3 cell line overexpresses the HER2/c-erb-2 gene product.

Complete Growth Medium	The base medium for this cell line is ATCC-formulated McCoy''''''''s 5a Medium Modified, Catalog No. 30-2007. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
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 which 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 is recommended Medium Renewal: 2 to 3 times per week
Cryopreservation	Freeze medium: Complete growth medium, 95%; DMSO, 5% Storage temperature: liquid nitrogen vapor temperature
Culture Conditions	Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C