Product Description

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Cell Biology
ATCC® Number: SCRC-1008™    Price: £158.00; €188.00
Designations: MEF (C57BL/6) [MEF-BL/6-1]
Depositors:  ATCC
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Mus musculus (mouse)
Morphology: fibroblast
Source: Strain: C57BL/6
Organ: embryo, whole
Cell Type: fibroblast
Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Isolation: United States
Isolation date: January 22, 2003
Age: 14 days gestation embryo
Gender: male and female mixed
Comments: The cell line was established by ATCCin 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. ATCChas 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).
Propagation: ATCC 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%

    • Atmosphere: air, 95%; carbon dioxide (CO2), 5%
    Temperature: 37.0°C
    Subculturing: Protocol: 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, ATCCcat# 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/0.53mM 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
    Preservation: Freeze medium: Complete growth medium, supplemented with an additional 40% FBS and 10% DMSO(v/v)
    Storage temperature: liquid nitrogen vapor phase
    Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
    recommended serum:ATCC 30-2020
    derivative:ATCC SCRC-1008.1
    derivative:ATCC SCRC-1008.2
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