This is a diploid human cell line with 46,XX karyotype. Polyploidy occurred at 6.9%chromosomes were normal. The chromosome N9 pair was heteromorphic for the centromeric heterochromatin having one with the normal size and the other about twice the size of the normal.
Derivation
Hs 578Bst was derived by A.J. Hackett, et al. from normal breast tissue peripheral to an infiltrating ductal carcinoma which was the source for Hs 578T (see ATCC HTB-126).
Hs 578Bst may have been myoepithelial in origin since the cells possessed microfilaments and clusters of pinocytotic vesicles similar to those seen in myoepithelia in vivo.
Receptor Expression
Epidermal growth factor (EGF)
Comments
No desmosomes were observed, estrogen receptor protein was not present, and no endogenous viruses were detected.
A frozen ampule at unknown population doubling (PDL) was received at the ATCC in 1983. Cells had the potential to reach approximay 22 more population doublings before the onset of senescence. See Batch Specific information for PDL of current Distribution freeze.
Complete Growth Medium
The base medium for this cell line is ATCC Hybri-Care Medium, Catalog No. 46-X. Hybri-Care Medium is supplied as a powder and should be reconstituted in 1 L cell-culture-grade water and supplemented with 1.5 g/L sodium bicarbonate. To make the complete growth medium, add the following components to the base medium: 30 ng/ml mouse EGF; fetal bovine serum to a final concentration of 10%.
Subculturing
Volumes used in this protocol are for 75 cm2 flasks; 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 0.25% (w/v) Trypsin-0.53mM 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 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.
Medium Renewal: 2 to 3 times per week
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
Complete growth medium, 95%; DMSO, 5%. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
