BALB SFME Serum Free Mouse Embryo (ATCC® CRL-9392)

Organism: Mus musculus, mouse  /  Cell Type: Astrocyte  /  Tissue: embryo  /  Disease: normal

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Organism Mus musculus, mouse
Tissue embryo
Cell Type Astrocyte
Product Format frozen
Morphology fibroblast
Culture Properties loosely adherent
Biosafety Level 1
Disease normal
Age embryo; 16 days
Strain BALB/c
Storage Conditions liquid nitrogen vapor phase
Karyotype modal number = 40
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Derivation
This neural stem cell line was derived from 16 day old BALB/c mouse embryos grown in serum-free medium.
Tumorigenic No, the cells were not tumorigenic in immunosuppressed mice.
Effects

Yes, the cells did form colonies in semisolid medium. 

No, the cells were not tumorigenic in immunosuppressed mice


Comments

BALB/c SFME cells infrequently form colonies in soft agar and are not tumorigenic if injected into nude mice.

Either serum or TGF-beta induce astrocyte differentiation accompanied by GFAP (glial fibrillary acidic protein) expression.

The presence of serum causes cell growth arrest. This process is reversible upon removal of the serum.

When cultured in serum-free medium, BALB SFME cells reportedly can be propagated for extended periods without undergoing crisis or gross chromosomal aberration.

Complete Growth Medium A 1:1 mixture of Dulbecco's Modified Eagle's Medium and Ham's F12 medium with 2.5 mM L-glutamine, 1.2 g/L sodium bicarbonate, 15mM HEPES and 0.5mM sodium puruvate supplemented with: 0.010 mg/ml bovine insulin 0.010 mg/ml human transferrin 1% chemically defined lipids (LifeTechnologies cat.# 11905-031) 50 ng/ml mouse epidermal growth factor 10 nM sodium selenite
Subculturing Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Note: Coat culture vessels with 0.01 mg/mL fibronectin solution before adding cells. Add culture medium plus an equal volume of 0.01 mg/mL fibronectin solution sufficient to cover the flask surface; immediately mix by rocking (Do not premix medium with fibronectin). Incubate at 37°C for at least 30 minutes. Suction to remove excess liquid prior to adding cells. Pre-coated culture vessels can be stored with medium for several days in the incubator.
  1. Attached cells may be removed with 0.05% trypsin - 0.53mM EDTA. The cells are very sensitive to trypsin; therefore the trypsinization process should take only a few seconds.
  2. Immediately inhibit the trypsin with equal volume of cold soybean trypsin inhibitor (0.1%) and dilute the cell suspension 10 fold with cold medium.  
  3. Pellet cells, resuspend in fresh medium and dispense into new pre-coated flasks. Non-attached cells can be spun down and replated in pre-coated flasks.
  4. Initially plate cells with half volume of medium in 5% CO2 in air atmosphere, wait 2-3 hours, and then add additional volume of medium.
    Note: The cells are very density and temperature sensitive. Confluencies above 80% will cause cells to form clumps and detach.  

DO NOT LEAVE FLASKS AT ROOM TEMPERATURE OR CELLS WILL DETACH.


Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:10 is recommended
Medium Renewal: Three to four times weekly with pre-warmed medium
Cryopreservation
Complete growth medium supplemented with 5% (v/v) DMSO. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
Note: The cells are very density and temperature sensitive. Confluencies above 80% will cause cells to form clumps and detach. Omission of EGF (epidermal growth factor) causes the cells to undergo apoptosis. Cells will detach at room temperature.
Name of Depositor Oregon State University
References

Loo DT, et al. Extended culture of mouse embryo cells without senescence: inhibition by serum. Science 236: 200-202, 1987. PubMed: 3494308

Rawson C, et al. Death of serum-free mouse embryo cells caused by epidermal growth factor deprivation is prevented by cycloheximide, 12-O-tetradecanoylphorbol-13-acetate, or vanadate. Exp. Cell Res. 186: 177-181, 1990. PubMed: 2153551

Rawson CL, et al. Death of serum-free mouse embryo cells caused by epidermal growth factor deprivation. J. Cell Biol. 113: 671-680, 1991. PubMed: 2016341

Loo D, et al. Serum-free mouse embryo cells: growth responses in vitro. J. Cell. Physiol. 139: 484-491, 1989. PubMed: 2786879

Ernst T. Karyotypic stability of serum-free mouse embryo (SFME) cells. Cytotechnology 5: 211-222, 1991. PubMed: 1367375

Rawson C, et al. Serum inhibition of proliferation of serum-free mouse embryo cells. Exp. Cell Res. 192: 271-277, 1991. PubMed: 1898591

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
References

Loo DT, et al. Extended culture of mouse embryo cells without senescence: inhibition by serum. Science 236: 200-202, 1987. PubMed: 3494308

Rawson C, et al. Death of serum-free mouse embryo cells caused by epidermal growth factor deprivation is prevented by cycloheximide, 12-O-tetradecanoylphorbol-13-acetate, or vanadate. Exp. Cell Res. 186: 177-181, 1990. PubMed: 2153551

Rawson CL, et al. Death of serum-free mouse embryo cells caused by epidermal growth factor deprivation. J. Cell Biol. 113: 671-680, 1991. PubMed: 2016341

Loo D, et al. Serum-free mouse embryo cells: growth responses in vitro. J. Cell. Physiol. 139: 484-491, 1989. PubMed: 2786879

Ernst T. Karyotypic stability of serum-free mouse embryo (SFME) cells. Cytotechnology 5: 211-222, 1991. PubMed: 1367375

Rawson C, et al. Serum inhibition of proliferation of serum-free mouse embryo cells. Exp. Cell Res. 192: 271-277, 1991. PubMed: 1898591

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm