184A1 (ATCC® CRL-8798)

Organism: Homo sapiens, human  /  Cell Type: chemically transformed  /  Tissue: mammary gland; breast/epithelium  / 

Organism Homo sapiens, human
Tissue
mammary gland; breast/epithelium
Cell Type chemically transformed
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Age 21 years
Gender female
Karyotype 45, XX
Derivation
Cells derived from the tissue were exposed to benzo(a)pyrene, and a transformed line was established.
The 184A1 cell line was established from normal mammary tissue obtained from a normal reduction mammoplasty.
Clinical Data
female
21 years
Comments The line appears to be immortal, but is not malignant.

When seeded at low density, the cells grow as single cells.
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):
  • 0.005 mg/ml transferrin
  • 1 ng/ml cholera toxin
Note: Do not filter complete medium
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.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. To remove trypsin-EDTA solution, 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 serum-free growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
  6. Place culture vessels in incubators at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:6 is recommended
Medium Renewal: Every 2 to 3 days
Cryopreservation
Culture medium, 75%; fetal bovine serum, 15%; glycerol, 10%
Culture Conditions
Temperature: 37°C
STR Profile
Amelogenin: X
CSF1PO: 10,11
D13S317: 11
D16S539: 11,12
D5S818: 11,13
D7S820: 9,11
THO1: 9.3
TPOX: 11
vWA: 18,19
Name of Depositor The United States of America
References

Milo G, et al. (eds.)., Transformation of human epithelial cells. Boca Raton, FL: CRC Press; 1992.

Stampfer MR. Continuous human cell lines and method of making same. US Patent 4,808,532 dated Feb 28 1989

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

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. PubMed: 3857588

Basic Documentation
References

Milo G, et al. (eds.)., Transformation of human epithelial cells. Boca Raton, FL: CRC Press; 1992.

Stampfer MR. Continuous human cell lines and method of making same. US Patent 4,808,532 dated Feb 28 1989

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

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. PubMed: 3857588