| BCRJ Code | 0386 |
| Cell Line | HCC70 |
| Species | Homo sapiens |
| Vulgar Name | Human |
| Tissue | Breast; mammary gland/duct |
| Cell Type | Epithelial |
| Morphology | Epithelial |
| Disease | TNM stage IIIA, grade 3, primary ductal carcinoma |
| Growth Properties | Adherent, The line grows as attached medium-sized epithelial cells |
| Sex | Female |
| Age/Ethinicity | 49 Year / |
| Derivation | The HCC70 cell line was initiated from a primary ductal carcinoma on June 3, 1992, and took 44 months to establish. |
| Products | Epithelial glycoprotein 2 [EGP2]; cytokeratin 19 |
| Biosafety | 1 |
| Addtional Info | The cells are poorly differentiated. The tumor was classified as TNM Stage IIIA, grade 3, invasive ductal carcinoma with metastases in 4 out of 17 lymph nodes. The cells overexpress p53, and are negative for the expression of Her2-neu oncogenes. HCC70 is positive for the epithelial cell specific marker Epithelial Glycoprotein 2 (EGP2) and for cytokeratin 19. |
| Culture Medium | RPMI-1640 medium modified to contain 2 mM L-glutamine, 4500 mg/L glucose with 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 1.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until the 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. Transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels. Maintain cultures at a cell concentration between 4x104 and 4 x 105 cells/cm2. Place culture vessels in incubators 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 | Every 2 to 3 days |
| Subculturing Subcultivation Ratio | 1:4 to 1:6 |
| 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 | Kao J, et al. Molecular profiling of breast cancer cell lines defines relevant tumor models and provides a resource for cancer gene discovery. PlosONE 4 (7): e 6146, 2009 Kim MS, et al. Breast cancer diagnosis using a microfluidic multiplexed immunohistochemistry platform. PlosONE 5 (5): e10441, 2010 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 | Leticia Batista Azevedo Rangel - Universidade Federal do Espírito Santo |
| Cellosaurus | CVCL_1270 |
