| BCRJ Code | 0280 |
| Cell Line | HCC1954 |
| Species | Homo sapiens |
| Vulgar Name | Human |
| Tissue | Mammary Gland; Breast/Duct |
| Cell Type | Epithelial |
| Morphology | Large Epithelial With Occasional Vacuoles |
| Disease | Tnm Stage Iia, Grade 3, Ductal Carcinoma |
| Growth Properties | Adherent |
| Sex | Female |
| Age/Ethinicity | 61 Year / |
| Derivation | HCC1954 was derived from a primary stage IIA, grade 3 invasive ductal carcinoma with no lymph node metastases. The HCC1954 is a poorly differentiated cell line initiated on October 30, 1995; it took about 4 months to establish. |
| DNA Profile | Amelogenin: X CSF1PO: 10 D13S317: 8,9 D16S539: 9,11 D5S818: 11 D7S820: 10,11 THO1: 6,7 TPOX: 8,9 vWA: 18,19 |
| Products | Epithelial glycoprotein 2 [EGP2]; cytokeratin 19 |
| Biosafety | 1 |
| Addtional Info | HCC1954 is positive for the epithelial cell specific marker Epithelial Glycoprotein 2 and for cytokeratin 19, and is negative for expression of estrogen receptor (ER) and progesterone receptor (PR). Her2/neu is overexpressed in the ELISA assay |
| Culture Medium | RPMI-1640 medium modified to contain 2 mM L-glutamine, 4500 mg/L glucose fetal bovine serum to a final concentration of 10%. |
| Subculturing | Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Remove and discard culture medium. Briefly rinse the cell layer with PBS without calcium and magnesium 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. NOTE: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 12 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 6th edition, published by Alan R. Liss, N.Y., 2010. |
| Subculturing Medium Renewal | 2 to 3 times a week |
| Subculturing Subcultivation Ratio | 1:4 to 1:8 |
| Culture Conditions | Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C |
| Cryopreservation | 95% FBS + 5% DMSO (Dimethyl sulfoxide) |
| Thawing Frozen Cells | SAFETY PRECAUTION:
It is strongly recommended to always wear protective gloves, clothing, and a full-face mask when handling frozen vials. Some vials may leak when submerged in liquid nitrogen, allowing nitrogen to slowly enter the vial. Upon thawing, the conversion of liquid nitrogen back to its gas phase may cause the vial to explode or eject its cap with significant force, creating flying debris.
NOTE: It is important to avoid excessive alkalinity of the medium during cell recovery. To minimize this risk, it is recommended to place the culture vessel containing the growth medium in the incubator for at least 15 minutes before adding the vial contents. This allows the medium to stabilize at its normal pH (7.0 to 7.6). |
| References | 38266: Gazdar AF, et al. Characterization of paired tumor and non-tumor cell lines established from patients with breast cancer. Int. J. Cancer 78: 766-774, 1998. PubMed: 9833771 |
| Depositors | DENISE DE ABREVE PEREIRA; UNIVERSIDADE FEDERAL DO RIO DE JANEIRO |
| Cellosaurus | CVCL_1259 |
