MEF (C57BL/6) [MEF-BL/6-1] cell line小鼠胚胎成纤维细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心
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- 货 号:MEF (C57BL/6) [MEF-BL/6-1]
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MEF (C57BL/6) [MEF-BL/6-1] cell line小鼠胚胎成纤维细胞株
MEF (C57BL/6) [MEF-BL/6-1]
SCRC-1008™
Product category
Animal cells
Organism
Mus musculus, mouse
Classification
Eukaryota, Animalia, Metazoa, Chordata, Vertebrata, Tetrapod
Cell type
fibroblast
Morphology
Fibroblast
Tissue
Embryo
Applications
Stem cell research
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).
Characteristics
Growth properties
Adherent
Derivation
The cell line was established by ATCC in 2003 from dissociated C57BL/6 mouse embryos.
Age
14 days gestation
Gender
Male and female mixed
Strain
C57BL/6
Handling information
Unpacking and storage instructions
Check all containers for leakage or breakage.
Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below _x001f_-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete 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).
Temperature
37°C
Atmosphere
95% Air, 5% CO2
Handling procedure
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 completes DMEM to a 15 mL centrifuge tube. Use an additional 1 mL of media to rinse the vial and transfer the liquid to the 15 mL tube. Add 4 mL of complete DMEM to bring the total volume to 10 mL.
Gently mix and pellet the cells by centrifugation at 270 x g for 5 minutes.
Discard the supernatant and resuspend the cells with 10 mL fresh growth medium (warm) and plate the cells at seed density of 1X104 cells/cm2.
Add fresh growth medium (warm) to the appropriate size 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 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.
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 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.
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.
Transfer all cells into a centrifuge bottle or tube and centrifuge at 270 x g for 5 minutes.
Remove and discard the supernatant
Add 10 mL complete growth medium to the cell pellet and with 10 mL pipette resuspend the cells gently (create a single-cell suspension).
Add more complete growth medium (Table 1) to the cell suspension as needed to plate cells at approximately 0.8 X 104 cells/cm2.
Place flasks in the incubator @ 37°C with a 5% CO2 in air atmosphere
Flask/Plate
Growth Area (cm2)
1xPBS (mL)
Trypsin/EDTA (mL)
Equal vol. Complete Growth Medium (mL)
Growth Medium (mL)
T225
225
10 ± 0.2
6 ± 0.2
6 ± 0.2
30
75
75
5 ± 0.1
3 ± 0.1
3 ± 0. 1
12
T25
25
3 ± 0.1
1.5 ± 0.1
1.5 ± 0.1
6
6 well
9.5
1 ± 0.1
1 ± 0.1
1 ± 0.1
3
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 5th edition, published by Alan R. Liss, N.Y., 2005.
Subcultivation Ratio: 1:5 to 1:8
Reagents for cryopreservation
Complete growth medium supplemented with 40% (v/v) FBS and 10% (v/v) DMSO
Supplier来源:BioVector NTCC Inc.
TEL电话:400-800-2947
Website网址: http://www.biovector.net
MEF (C57BL/6) [MEF-BL/6-1]
SCRC-1008™
Product category
Animal cells
Organism
Mus musculus, mouse
Classification
Eukaryota, Animalia, Metazoa, Chordata, Vertebrata, Tetrapod
Cell type
fibroblast
Morphology
Fibroblast
Tissue
Embryo
Applications
Stem cell research
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).
Characteristics
Growth properties
Adherent
Derivation
The cell line was established by ATCC in 2003 from dissociated C57BL/6 mouse embryos.
Age
14 days gestation
Gender
Male and female mixed
Strain
C57BL/6
Handling information
Unpacking and storage instructions
Check all containers for leakage or breakage.
Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below _x001f_-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete 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).
Temperature
37°C
Atmosphere
95% Air, 5% CO2
Handling procedure
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 completes DMEM to a 15 mL centrifuge tube. Use an additional 1 mL of media to rinse the vial and transfer the liquid to the 15 mL tube. Add 4 mL of complete DMEM to bring the total volume to 10 mL.
Gently mix and pellet the cells by centrifugation at 270 x g for 5 minutes.
Discard the supernatant and resuspend the cells with 10 mL fresh growth medium (warm) and plate the cells at seed density of 1X104 cells/cm2.
Add fresh growth medium (warm) to the appropriate size 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 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.
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 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.
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.
Transfer all cells into a centrifuge bottle or tube and centrifuge at 270 x g for 5 minutes.
Remove and discard the supernatant
Add 10 mL complete growth medium to the cell pellet and with 10 mL pipette resuspend the cells gently (create a single-cell suspension).
Add more complete growth medium (Table 1) to the cell suspension as needed to plate cells at approximately 0.8 X 104 cells/cm2.
Place flasks in the incubator @ 37°C with a 5% CO2 in air atmosphere
Flask/Plate
Growth Area (cm2)
1xPBS (mL)
Trypsin/EDTA (mL)
Equal vol. Complete Growth Medium (mL)
Growth Medium (mL)
T225
225
10 ± 0.2
6 ± 0.2
6 ± 0.2
30
75
75
5 ± 0.1
3 ± 0.1
3 ± 0. 1
12
T25
25
3 ± 0.1
1.5 ± 0.1
1.5 ± 0.1
6
6 well
9.5
1 ± 0.1
1 ± 0.1
1 ± 0.1
3
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 5th edition, published by Alan R. Liss, N.Y., 2005.
Subcultivation Ratio: 1:5 to 1:8
Reagents for cryopreservation
Complete growth medium supplemented with 40% (v/v) FBS and 10% (v/v) DMSO
Supplier来源:BioVector NTCC Inc.
TEL电话:400-800-2947
Website网址: http://www.biovector.net
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