MEF (C57BL/6) [MEF-BL/6-1] (ATCC® SCRC-1008)

Organism: Mus musculus, mouse  /  Cell Type: Fibroblast  /  Tissue: Embryo  / 

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Organism Mus musculus, mouse
Tissue
Embryo
Cell Type Fibroblast
Product Format frozen
Morphology Fibroblast
Culture Properties Adherent
Biosafety Level 1

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.

Age 14 days gestation embryo
Gender male and female mixed
Strain C57BL/6
Applications
The cells can be used as a feeder layer to support the growth of embryonic stem (ES) cells and for the maintenance of ES cells in the undifferentiated state. The growth of these cells must be arrested before they can be used as a feeder layer. ATCC has successfully treated the cells with mitomycin C for use as a feeder layer. If the MEFs are being used as a feeder layer for ES cells, it is not recommended to use them past passage no. 6 (P6).
Storage Conditions liquid nitrogen vapor phase
Derivation
The cell line was established by ATCC in 2003 from dissociated C57BL/6 mouse embryos.
Clinical Data
male and female mixed
Comments
The cell line was established by ATCC in 2003 from dissociated C57BL/6 mouse embryos. The cells can be used as a feeder layer to support the growth of embryonic stem (ES) cells and for the maintenance of ES cells in the undifferentiated state. The growth of these cells must be arrested before they can be used as a feeder layer. ATCC has successfully treated the cells with mitomycin C for use as a feeder layer. If the MEFs are being used as a feeder layer for ES cells, it is not recommended to use them past passage no. 6 (P6).
Complete Growth Medium The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium:
  • fetal bovine serum to a final concentration of 15%

  • This medium is formulated for use with a 5% CO2 in air atmosphere. (Standard DMEM formulations contain 3.7 g/L sodium bicarbonate and a 10% CO2 in air atmosphere is then recommended).
    Subculturing
    Volumes used in this protocol are for 75 cm2 flask; 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 1X PBS (Ca+2/Mg+2-free, ATCC cat# SCRR-2201) to remove all traces of serum that contains trypsin inhibitor.
    3. Add 2.0 to 3.0 mL of 0.25% (w/v) Trypsin/0.53 mM EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 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.
    6. Incubate cultures at 37°C.
    Subcultivation Ratio: 1:5 to 1:8
    Cryopreservation
    Liquid nitrogen vapor phase
    Culture Conditions
    Atmosphere: air, 95%; carbon dioxide (CO2), 5%
    Temperature: 37.0°C
    Name of Depositor ATCC
    Year of Origin January 22, 2003
    Notice: Necessary PermitsPermits

    These permits may be required for shipping this product:

    • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
    Basic Documentation