| 產(chǎn)品名稱 |
HCC1569 |
| 商品貨號 |
B164561 |
| Organism |
Homo sapiens, human |
| Tissue |
mammary gland; breast |
| Cell Type |
Epithelial |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
mixed adherent-suspension |
| 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 |
TNM stage IV, grade 3, primary metaplastic carcinoma |
| Age |
70 years |
| Gender |
female |
| Ethnicity |
Black |
| Karyotype |
polyploid |
| Derivation |
The cell line is derived from an older patient with a germline mutation in the FHIT gene. The FHIT gene is located at 3p14.2 and the mutation is a transversion at nucleotide 651 (G to T).The patient's daughter carries the same gene mutation. This cell line was initiated on 3/8/95 and took 19 months to establish. |
| Clinical Data |
70 years
Black
female
The patient received prior chemotherapy and had no family history of breast cancer.
The tumor was classified as TNM stage IV, grade 3, metaplastic carcinoma with 4 out of 18 lymph node metastasis. |
| Receptor Expression |
estrogen receptor, negative progesterone receptor, negative |
| Oncogene |
her2/neu +, p53 - |
| Genes Expressed |
Epithelial glycoprotein 2 [EGP2]; cytokeratin 19 |
| Cellular Products |
Epithelial glycoprotein 2 [EGP2]; cytokeratin 19 |
| Comments |
The patient received prior chemotherapy and had no family history of breast cancer. The tumor was classified as TNM stage IV, grade 3, metaplastic carcinoma with 4 out of 18 lymph node metastasis.
The cells are poorly differentiated.
The cells are positive for expression of Her2-neu and negative for expression of p53.
HCC1569 is positive for the epithelial cell specific marker Epithelial Glycoprotein 2 (EGP2) and for cytokeratin 19.
The cells are negative for expression of estrogen receptor (ER) and for expression of progesterone receptor (PR) by immunohistochemistry, but PR expression was detectable at a low level by cytosolic protein assay. |
| Complete Growth Medium |
The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, ATCC 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (ATCC 30-2020) to a final concentration of 10%.
|
| Subculturing |
Volumes used in this protocol are for a 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
- Remove culture medium, which contains suspended cells, to a centrifuge tube.
- Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 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 10 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.
- To remove trypsin-EDTA solution, transfer cell suspension to the centrifuge tube containing the medium and cells from step #1 and centrifuge 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.
- Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended
Medium Renewal: Every 2 to 3 days.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Animal Cells: A Manual Of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005. |
| Cryopreservation |
culture medium 95%; DMSO, 5% |
| Culture Conditions |
Temperature: 37°C Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
| Name of Depositor |
AF Gazdar, AK Virmani |
| References |
Ahmadian M, et al. Analysis of the FHIT gene and FRA3B region in sporadic breast cancer, preneoplastic lesions, and familial breast cancer probands. Cancer Res. 57: 3664-3668, 1997. PubMed: 9288768
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
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