EO771 cell line小鼠髓样乳腺癌贴壁细胞-BioVector NTCC典型培养物保藏中心

EO771 Cell Line小鼠髓样乳腺癌细胞株

Cat No.: BioVector963461

Product category

Animal cells

Organism

Mus musculus, mouse

Morphology

epithelial-like

Tissue

Breast; Mammary gland

Disease

Carcinoma; Breast

Applications

3D cell culture

Specific applications

This cell line can be used to analyze genes that regulate metastasis of breast cancer and/or drugs that might reduce spontaneous metastasis in immune competent C57Bl/6 mice.

Characteristics

Cells per vial

Approximately 2.0 to 3.0 x 106

Volume

1.0 mL

Growth properties

Adherent

Comments

The cells display an elongated morphology. Depositor note: cells are genetically unstable in continuous culture. The EO771 mammary tumor was first reported in 1948 as a spontaneous mammary carcinoma arising in a C57Bl/6 mouse.

Images:

Complete medium

The base medium for this cell line is Dulbecco's Modified Eagle's Medium (DMEM; BioVector 30-2002). To make the complete medium add the following components to the base medium:

· 10% Fetal Bovine Serum (FBS; BioVector 30-2020)

· 20 mM HEPES (Thermo Fisher Scientific cat# 15630-080)

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 -130°C, preferably in liquid nitrogen vapor, until ready for use.

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 6.0 x 104 and 8.0 x 104 viable cells/cm2.

6. Incubate cultures at 37°C.

Interval: Maintain cultures at a cell concentration between 5.0 X 104 and 1.0 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

DMEM + 7.5% FBS + 5% DMSO

Quality control specifications

Bacterial and fungal testing

Not detected

Mycoplasma contamination

Not detected

Virus testing

Hepatitis B virus (HBV): Not detected

Cytomegalovirus (CMV): Not detected

Human Immunodeficiency virus (HIV): Not detected

Epstein-Barr virus (EBV): Not detected

Human papillomavirus (HPV): Not detected

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