MCF-7 Tam1 human Breast Cancer tamoxifen-resistant Cell Line BioVector NTCC质粒载体菌种细胞基因保藏中心

MCF-7 Tam1 human Breast Cancer tamoxifen-resistant Cell Line

MCF-7 TamoxifenR are cells exhibiting epithelial morphology that were isolated from the breasts of a female patient with ductal carcinoma. This product is derived from MCF-7 (NTCC HTB-22) cell line, continuously cultured with 1 µM 4-OH-tamoxifen for 8-12 months to obtain a tamoxifen resistant cell line. This cell line has applications for cancer research.
Product category
Human cells
Organism
Homo sapiens, human
Morphology
epithelial-like
Tissue
Breast; Mammary gland
Disease
Carcinoma; Ductal
Applications
3D cell culture
Cancer research

Characteristics
Cells per vial
Approximately 2.0 to 3.0 x 106
Growth properties
Adherent
Gender
Female
Comments
Derived from MCF-7 (NTCC HTB-22) cell line, continuously cultured with 1 µM 4-OH-tamoxifen for 8-12 months to obtain a tamoxifen resistant cell line
Handling information
Unpacking and storage instructions
1.Check all containers for leakage or breakage.
2.Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below _x001f_-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete medium
The base medium for this cell line is Dulbecco's Minimum Essential Medium (DMEM; NTCC 30-2002). To make the complete medium add the following components to the base medium:
·10% Fetal Bovine Serum (FBS; NTCC 30-2020)
·10 µg/mL human insulin (Sigma cat# I9278)
·1 µM 4-hydroxytamoxifen

Temperature
37°C
Atmosphere
95% Air, 5% CO2
Handling procedure
To ensure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.  
1.Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water.  Thawing should be rapid (approximately 2 minutes).
2.Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
3.Transfer the vial contents to a centrifuge tube containing  9.0 mL complete culture medium. and spin at approximately 125 x g for 5 to 7 minutes.
4.Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). It is important to avoid excessive alkalinity of the medium during recovery of the cells.  It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6). pH (7.0 to 7.6).
5.Incubate the culture at 37°C in a suitable incubator.  A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.

Subculturing procedure
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
1.Remove and discard culture medium.
2.Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
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.Add appropriate aliquots of the cell suspension to new culture vessels.
Cultures can be established between 1.5 x 104 and 5.0 x 104 viable cells/cm2.
6.Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 1.0 X 104 and 3.1 X 105 cell/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
Reagents for cryopreservation
Culture medium (without 4-hydroxytamoxifen) + 5% DMSO (NTCC 4-X)
Quality control specifications
Bacterial and fungal testing
Not detected
Mycoplasma contamination
Not detected
Population doubling time
Approximately 1 to 2 weeks

Supplier来源:BioVector NTCC Inc.
TEL电话:400-800-2947
Website网址: http://www.biovector.net