MEF (CF-1)細胞

生長狀態(tài)： 貼壁生長

運輸方式： 凍存運輸

細胞形態(tài)： 成纖維樣

年限： 14 days gestation embryo

器官來源： 胚胎

細胞類型： 成纖維細胞

是否是腫瘤細胞： 0

物種來源： 小鼠

品系： CF-1

ATCC Number： SCRC-1040?

數(shù)量： 大量

Designations: MEF (CF-1)

Depositors: ATCC

MEF (CF-1)細胞Biosafety Level: 1

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Mus musculus

Morphology: fibroblast

Source: Organ: embryo

Strain: CF-1

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: Isolation date: 2003

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.

ATCC has successfully irradiated (SCRC-1040.1) and treated the cells with Mitomycin C (SCRC-1040.2a) for use as a feeder layer.

MEF (CF-1)細胞The cell line was established by ATCC in 2003 from embryonic day 14 (E14) CF-1 mouse embryos.

The growth of these cells should be arrested before being used 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)

Age: 14 days gestation embryo

Gender: male and female mixed

Comments: The cell line was established by ATCC in 2003 from embryonic day 14 (E14) CF-1 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 should be arrested before being used as a feeder layer. ATCC has successfully irradiated (SCRC-1040.1) and treated the cells with Mitomycin C (SCRC-1040.2a) 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: 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.

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.

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.

Transfer the vial s contents plus 5 ml of complete medium MEF (CF-1)細胞(see ATCC complete growth medium 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.

Gently mix and pellet the cells by centrifugation @ 270x g for 5 minutes.

Discard the supernatant and resuspend the cells with 10ml fresh growth medium (warm) and plate cells at seed density of 0.8 X 104 cells/cm2.

Add 5 ml more fresh growth medium (warm) to flask.

Incubate 37?C in a 5% CO2 in air atmosphere.

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).

Subculturing Procedure:

To insure the highest level of viability, be sure to warm media and Trypsin / EDTA to 37?C before using it on the cells. Cells should be split when they reach confluency. Split cells at approximately 0.4 X 104 cells/cm2

Remove and discard culture medium.

Briefly rinse the cell layer with 1XPBS (SCRR-2201) solution to remove all traces of serum, which contain trypsin inhibitor.

Add 5 ml of Trypsin-EDTA (0.25% (w/v) Trypsin-0.53 mM EDTA solution, ATCC # 30-2101) solution to flask and incubate for 1 minute, gently tapping the flask observe cells under an inverted microscope until cells detach (usually within 1 to 2 minutes).

Add 6.0 to 8.0 ml of complete growth medium and rinse surface of the flask to detach all cells. Gently pipetting up and down will break cell clumps.

Transfer all cells into a centrifuge bottle or tube and centrifuge at 271x g for 5 minutes.

Remove and discard the supernatant

Add 10 ml complete growth medium to cell pellet and with 10 ml pipette resuspend the cells gently (create a single-cell suspension).

Add more complete growth medium to cell suspension as needed to plate cells.

Place flasks in incubator @ 37°C with a 5% CO2 in air atmosphere.

Subcultivation Ratio: MEF (CF-1)細胞Plate the cells at approximately of 0.4 X 104 cells/cm2.

Medium Renewal: Twice a week or when pH decreases.

Preservation: Freeze medium: Complete growth medium supplemented with an additional 40% FBS and 10% DMSO

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-1040.1

derivative:ATCC SCRC-1040.2a

References: 89421: Nagy A, et al. Manipulating The Mouse Embryo: A Laboratory Manual. Third Edition: Cold Spring Harbor Press; 2003.