184B5細胞
ATCC Number： CRL-8799?
相關**： 正常
生長狀態： 貼壁生長
器官來源： **
組織來源： epithelium
數量： 大量
細胞類型： 其他細胞類型
是否是腫瘤細胞： 0
物種來源： 人
運輸方式： 凍存運輸
細胞形態： 上皮樣
年限： 21 years
184B5細胞Designations: 184B5
Depositors: The United States of America
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Homo sapiens
Morphology: epithelial
Source: Organ: mammary gland; breast
Tissue: epithelium
Disease: normal
Cell Type: chemically transformed
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Tumorigenic: No
Cytogenetic Analysis: 47, XX
Age: 21 years
Gender: female
Comments: 184B5細胞The 184B5 cell line was established from normal mammary tissue obtained from a normal reduction mammoplasty.
Cells derived from the tissue were exposed to benzo(a)pyrene, and a transformed line was established.
The line appears to be immortal, but is not malignant.
When seeded at low density, the cells grow in tightly packed colonies.
Propagation: ATCC complete growth medium: The base medium for this cell line (MEBM) along with the additives can be obtained from Lonza/Clonetics Corporation as a kit: MEGM, Kit Catalog No. CC-3150. ATCC does not use the GA-1000 (gentamycin-amphotericin B mix) provided with kit. To make the complete growth medium, you will need to add the following components to the kit (sold separately):
1 ng/ml cholera toxin
Note: Do not filter complete medium
Temperature: 37.0°C
Subculturing: Protocol:
Remove and discard culture medium.
Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution.
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.
To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 xg for 5 to10 minutes.Discard supernatant and resuspend cells in fresh serum-free growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
Place culture vessels in incubators at 37?C.
Subcultivation Ratio: 184B5細胞A subcultivation ratio of 1:2 to 1:6 is recommended
Medium Renewal: Every 2 to 3 days
Preservation: Freeze medium: Complete growth medium suplemented with 10% (v/v) fetal bovine serum and 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
References: 21894: . Transformation of human epithelial cells. Boca Raton, FL: CRC Press; 1992.
21926: Stampfer MR. Continuous human cell lines and method of making same. US Patent 4,808,532 dated Feb 28 1989
22413: Walen KH, Stampfer MR. Chromosome analyses of human mammary epithelial cells at stages of chemical-induced transformation progression to immortality. Cancer Genet. Cytogenet. 37: 249-261, 1989. PubMed: 2702624
23289: Stampfer MR, Bartley JC. Induction of transformation and continuous cell lines from normal human mammary epithelial cells after exposure to benzo[a]pyrene. Proc. Natl. Acad. Sci. USA 82: 2394-2398, 1985. 184B5細胞PubMed: 3857588
