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90.74
90.74
規(guī)格:
貨期:
編號:B231739
品牌:Mingzhoubio

標(biāo)準(zhǔn)菌株
定量菌液
DNA
RNA

規(guī)格:
凍干粉
斜面
甘油
平板


產(chǎn)品名稱 90.74
商品貨號 B231739
Organism Homo sapiens, human
Tissue kidney
Cell Type transformed with adenovirus 5 DNA
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 Cells contain Human adenovirus sequences

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 fetus
Applications
packaging cell line
Storage Conditions liquid nitrogen vapor phase
Disclosure This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC.
Derivation
This line is an amphotropic retrovirus packaging cell line derived from the human embryonic kidney line, 293 (ATCC CRL-1573). 293 cells were co-transfected with the pIK6.1MCVampac plasmid (ATCC 75483) encoding the gag and pol genes from ectropic MMLV and the envelope gene from the 4070A amphotropic MLV and MC1 neo [U.S. Pat. 5,858,740]. The line does not depend on continued G418 selection to maintain the packaging genome over time.
Comments
This line is an amphotropic retrovirus packaging cell line derived from the human embryonic kidney line, 293 (ATCC CRL-1573). 293 cells were co-transfected with the pIK6.1MCVampac plasmid (ATCC 75483) encoding the gag and pol genes from ectropic MMLV and the envelope gene from the 4070A amphotropic MLV and MC1 neo [U.S. Pat. 5,858,740]. The line does not depend on continued G418 selection to maintain the packaging genome over time.
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: bovine calf serum to a final concentration of 10%.
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.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 2 x 103 to 6 x 103 viable cells/cm2 is recommended. Maintain cultures at a cell concentration between 2 x 103 and 2.5 x 105 cells/cm2. 
    NOTE:  Coat vessels with 0.1% pocine gelatin for 30 minutes to enhance attachment.
  6. Incubate cultures at 37°C.

Subcultivation Ratio: 1:4 to 1:6
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation

Complete growth medium described above 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%
Population Doubling Time 22 hours
Name of Depositor Cell Genesys
U.S. Patent Number
References

Finer MH, et al. Method for production of high titer virus and high efficiency retroviral mediated transduction of mammalian cells. US Patent 5,858,740 dated Jan 12 1999

Finer MH, et al. Method for production of high titer virus and high efficiency retroviral mediated transduction of mammalian cells. US Patent 6,506,604 dated Jan 14 2003

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.

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