宁波泰斯拓生物

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浙江省宁波市镇海区庄市街道兴庄路9号创e慧谷42号楼B幢401室
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MEF (C57BL/6) [MEF-BL/6-1]

货号 TS162722
中文名称 null
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产品简介
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产品名称: MEF (C57BL/6) MEF-BL/6-1
商品货号: TS162722
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
Complete Growth Medium: The base medium for this cell line is ATCC-formulated Dulbeccos Modified Eagles 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: To insure the highest level of viability, be sure to warm media and Trypsin / EDTA to 37ºC before using it on the cells. Cells should be split when they reach confluency. A split base on seed density of 2 X 104 cells/cm2 is recommended.
    1. Remove and discard culture medium.
    2. Briefly rinse the cell layer with 1XPBS (SCRR-2201) solution to remove all traces of serum, which contain trypsin inhibitor.
    3. Add Trypsin-EDTA (0.25% Trypsin-0.53 mM EDTA solution, ATCC# 30-2101) solution to the flask (Table 1) and incubate for 2 minutes. Gently tapping the flask, observe cells under an inverted microscope. Cells usually detach in 2 to 3 minutes.
    4. Add an equal volume complete of the growth medium (Table 1) and rinse surface of the flask to detach all the cells. Gently pipetting up and down will break cell clumps.xa0
    5. Transfer all cells into a centrifuge bottle or tube and centrifuge at 270 x g for 5 minutes.
    6. Remove and discard the supernatant
    7. Add 10 mL complete growth medium to the cell pellet and with 10 mL pipette resuspend the cells gently (create a single-cell suspension).
    8. Add more complete growth medium (Table 1) to the cell suspension as needed to plate cells at approximately 0.8 X 104 cells/cm2.
    9. Place flasks in the incubator @ 37°C with a 5% CO2 in air atmospherexa0

    Flask/Platexa0

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    Cryopreservation:

    Growth Area (cm2)xa0

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    Culture Conditions:

    xa01xPBS (mL)

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    Name of Depositor:

    Trypsin/EDTA (mL)xa0

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    Year of Origin:

    Equal vol. Complete Growth Medium (mL)xa0

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    References:

    Growth Medium (mL)xa0

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