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ATCC Number：SCRC-1008.1?
年限：14 days gestation embryo
細胞形態(tài)：成纖維樣
器官來源：其他
數(shù)量：大量
品系：C57BL/6
運輸方式：凍存運輸
細胞類型：成纖維細胞
生長狀態(tài)：貼壁生長
是否是腫瘤細胞：0
物種來源：小鼠
規(guī)格：0.1ml Designations: MEF (C57BL/6) IRR
Depositors: ?ATCC
Biosafety Level:1
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism: Mus musculus
Morphology:fibroblast


Source: Organ: embryo, whole
Strain: C57BL/6
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.
Applications:It is recommended that the feeder cells be plated 24 hours before use at 4 to 5 X 10(6) cells per T75 flask in order to obtain a supportive monolayer for stem cell growth.
These cells are provided for use as feeder cells to support the growth of stem cells in the undifferentiated state.
Age: 14 days gestation embryo
Gender: male and female mixed
Comments:These cells are provided for use as feeder cells to support the growth of stem cells in the undifferentiated state. They have been irradiated with 10,000 rads and will not replicate. The cells will begin to deteriorate in 7 to 10 days after plating. Once the feeder cells have attached, the culture medium can be changed to accommodate the cells to be supported. It is recommended that the feeder cells be plated 24 hours before use at 4 to 5 X 10(6) cells per T75 flask in order to obtain a supportive monolayer for stem cell growth.
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: Establishing and maintaining your culture: To insure the highest level of viability, be sure to warm media to 37?C before using it on the cells. Flasks do not need to be coated before plating MEFs.

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.
2. Remove the vial from the water bath as soon as the contents are half way thawed (approximately 90 seconds), 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 s contents plus 5 ml of complete medium (see below for recipe) to a 15 ml centrifuge tube. Use an additional 1 ml of medium to rinse the vial and transfer the liquid to the 15 ml tube. Add 4 ml of complete medium to bring the total volume to 10 ml.
4. Gently mix and pellet the cells by centrifugation @ 270x g for 5 minutes.
5. Discard the supernatant and resuspend the cells with 10ml fresh growth medium (warm) and transfer to one T75 flask.
6. Add 5 ml more fresh growth medium (warm) to flask.
7. Incubate 37?C in a 5% CO2 in air atmosphere.
8. Fluid change twice a week or when pH decreases. 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 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).

Cells should be plated 24 hours before use as a feeder layer for ES cells and kept for no more than 7 days
Medium Renewal: Twice a week or when pH decreases
Preservation: 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
source culture:ATCC SCRC-1008