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EpH4-Ev
EpH4-Ev
规格:
货期:
编号:TS178637
品牌:Testobio
产品名称: EpH4-Ev
商品货号: TS178637
Organism: Mus musculus, mouse
Tissue: breast epithelium (mammary gland)
Cell Type: epithelial cell
Product Format: frozen
Morphology: epithelial-like
Culture Properties: adherent
Biosafety Level: 2 Cells contain CMV and SV40 viral DNA sequences

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Gender: female
Applications:

This cell line can be used to study normal epithelial cell function and cell transformation.

Storage Conditions: liquid nitrogen vapor phase
Images: CRL-3063 Micrograph
Derivation: EpH4-Ev was produced by transfecting parental EpH4, a mouse mammary epithelial cell line, with an empty expression vector carrying a puromycin resistance gene.xa0 Stable clones were isolated by selection in medium containing 1.2 µg/mL puromycin.
Genes Expressed: puromycin resistance gene, expressed
Tumorigenic: No
Comments:

EpH4-Ev was produced by transfecting parental EpH4, an immortalized mouse mammary epithelial cell line, with an empty expression vector carrying a puromycin resistance gene. xa0Stable clones were isolated by selection in medium containing 1.2 mg/mL puromycin.

The EpH4 cell line presents an attractive model in which to study transformation because the cells exhibit morphological and functional characteristics typical of normal mammary epithelial cells. This cell line has been used to generate constitutively activated MEK1 transformed cell line B-MEKDD 116 (ATCC CRL-3069). There are three additional related cell lines: EpH 1424 (ATCC CRL-3071), EpH4 1424.1 (ATCC CRL-3209), EpH4 1424.2 (ATCC CRL-3210).

Complete Growth Medium: The base medium for this cell line is ATCC-formulated DMEM Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium:
  • 10% Bovine Calf Serum
  • 1.2 mcg/mL Puromycin

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.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbeccos phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 1.0 to 2.0 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. An inoculum of 4 x 103 to 8 x 103 viable cells/cm2 is recommended.
  6. Incubate cultures at 37°C. Subculture when the cell concentration is between 8 x 104 to 1.5 x 105 cells/cm2.

Subcultivation ratio: A subcultivation ratio of 1:6 to 1:15 is recommended.
Medium renewal: Every 2 to 3 days
Cryopreservation: Freeze medium: complete growth medium supplemented with an additional 10% bovine calf serum and 10% DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions: Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Name of Depositor: P Leder
References:

Fantin VR, et al. A novel mitochondriotoxic small molecule that selectively inhibits tumor cell growth. Cancer Cell 2(1): 29-42, 2002 PubMed: 12150823

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EpH4-Ev

  • 货号: TS178637
  • 好评
询价
  • 品牌 : TESTOBIO
产品名称: EpH4-Ev
商品货号: TS178637
Organism: Mus musculus, mouse
Tissue: breast epithelium (mammary gland)
Cell Type: epithelial cell
Product Format: frozen
Morphology: epithelial-like
Culture Properties: adherent
Biosafety Level: 2 Cells contain CMV and SV40 viral DNA sequences

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Gender: female
Applications:

This cell line can be used to study normal epithelial cell function and cell transformation.

Storage Conditions: liquid nitrogen vapor phase
Images: CRL-3063 Micrograph
Derivation: EpH4-Ev was produced by transfecting parental EpH4, a mouse mammary epithelial cell line, with an empty expression vector carrying a puromycin resistance gene.xa0 Stable clones were isolated by selection in medium containing 1.2 µg/mL puromycin.
Genes Expressed: puromycin resistance gene, expressed
Tumorigenic: No
Comments:

EpH4-Ev was produced by transfecting parental EpH4, an immortalized mouse mammary epithelial cell line, with an empty expression vector carrying a puromycin resistance gene. xa0Stable clones were isolated by selection in medium containing 1.2 mg/mL puromycin.

The EpH4 cell line presents an attractive model in which to study transformation because the cells exhibit morphological and functional characteristics typical of normal mammary epithelial cells. This cell line has been used to generate constitutively activated MEK1 transformed cell line B-MEKDD 116 (ATCC CRL-3069). There are three additional related cell lines: EpH 1424 (ATCC CRL-3071), EpH4 1424.1 (ATCC CRL-3209), EpH4 1424.2 (ATCC CRL-3210).

Complete Growth Medium: The base medium for this cell line is ATCC-formulated DMEM Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium:
  • 10% Bovine Calf Serum
  • 1.2 mcg/mL Puromycin

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.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbeccos phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 1.0 to 2.0 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. An inoculum of 4 x 103 to 8 x 103 viable cells/cm2 is recommended.
  6. Incubate cultures at 37°C. Subculture when the cell concentration is between 8 x 104 to 1.5 x 105 cells/cm2.

Subcultivation ratio: A subcultivation ratio of 1:6 to 1:15 is recommended.
Medium renewal: Every 2 to 3 days
Cryopreservation: Freeze medium: complete growth medium supplemented with an additional 10% bovine calf serum and 10% DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions: Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Name of Depositor: P Leder
References:

Fantin VR, et al. A novel mitochondriotoxic small molecule that selectively inhibits tumor cell growth. Cancer Cell 2(1): 29-42, 2002 PubMed: 12150823

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