ELT3 cell line大鼠子宫肌瘤细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

ELT3 cell line大鼠子宫肌瘤细胞株
Eker Leiomyoma Tumor-3 (ELT3) Cell Line

Cat No.: BioVector-CVCL_4616

Product category Animal cells

Organism  Rattus norvegicus, rat

Morphology  epithelial-like

Tissue  Uterus

Disease  Leiomyoma

Applications   3D cell culture

Characteristics

Cells per vial

Approximately 2.0 to 5.0 x 106

Volume

1.0 mL

Growth properties

Adherent

Gender

Female

Strain

Long Evans

Comments

Isolated from tumor from Eker rats carrying a germline defect in the Tsc2 gene (TscEk/+).

Diploid karyotype.

Loss of TSC2 tumor suppressor gene, expression of IGF-1, PDGF, and TGF-alpha.

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.

Complete medium

The complete medium for this cell line is DF8 medium:

The base medium is DMEM: F12. To make the complete medium add the following components to 500 mL of the base medium:

56 mL fetal bovine serum

1.6 µM ferrous sulfate

50 nM sodium selenite

12 nM vasopressin

10 nM cholesterol

0.2 μM hydrocortisone

10 µg/mL transferrin

nM T3

25 µg/mL insulin

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.

Quality control specifications

Bacterial and fungal testing: Not detected

Mycoplasma contamination: Not detected

Viability: ≥ 50%

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 3.0 x 104 viable cells/cm2.

6.Incubate cultures at 37°C.

Interval: Maintain cultures at a cell concentration between 1.0 X 104 and 8.0 X 104 cell/cm2.

Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended

Medium Renewal: 2 to 3 times per week

Supplier来源:BioVector NTCC Inc.
Website网址: http://www.biovector.net