As4.1 (ATCC® CRL-2193)

Organism: Mus musculus, mouse  /  Tissue: kidney  / 

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Organism Mus musculus, mouse
Tissue kidney
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2  [Cells contain papovavirus]
Gender female
Applications
transfection host
Storage Conditions liquid nitrogen vapor phase
Derivation
As4.1 was established from the ascites fluid of a six month old transgenic mouse harboring an intraparenchymal kidney tumor induced by transgene targeted tumorigenesis.
These transgenic mice were constructed with the renin T antigen fusion gene, pR2d4.6TAG which contains a transcriptional fusion between Ren-2d 5' flanking sequences and the structural gene for SV40 T antigen.
Clinical Data
female
Genes Expressed
renin
Cellular Products
renin
Comments The cell line was cloned by picking a small cluster from a dish seeded at low density.
The As4.1 cells express high levels of correctly processed renin mRNA from the endogenous Ren-1c locus.
They constitutively secrete prorenin and appear to store a proteolytically cleaved form of active renin intracellularly in storage granules/vesicles.
It has been reported that each As4.1 cell contains 1000 to 2000 copies of renin mRNA.
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 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.
  6. Incubate cultures at 37°C.

Subcultivation Ratio: 1:6 to 1:12
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%
Name of Depositor KW Gross
References

Jones CA, et al. Expression of murine renin genes during fetal development. Mol. Endocrinol. 4: 375-383, 1990. PubMed: 2188116

Sigmund CD, et al. Tissue and cell specific expression of a renin promoter-reporter gene construct in transgenic mice. Biochem. Biophys. Res. Commun. 170: 344-350, 1990. PubMed: 1695507

Sigmund CD, et al. Isolation and characterization of renin-expressing cell lines from transgenic mice containing a renin-promoter viral oncogene fusion construct. J. Biol. Chem. 265: 19916-19922, 1990. PubMed: 2174057

Petrovic N, et al. Role of proximal promoter elements in regulation of renin gene transcription. J. Biol. Chem. 271: 22499-22505, 1996. PubMed: 8798416

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
References

Jones CA, et al. Expression of murine renin genes during fetal development. Mol. Endocrinol. 4: 375-383, 1990. PubMed: 2188116

Sigmund CD, et al. Tissue and cell specific expression of a renin promoter-reporter gene construct in transgenic mice. Biochem. Biophys. Res. Commun. 170: 344-350, 1990. PubMed: 1695507

Sigmund CD, et al. Isolation and characterization of renin-expressing cell lines from transgenic mice containing a renin-promoter viral oncogene fusion construct. J. Biol. Chem. 265: 19916-19922, 1990. PubMed: 2174057

Petrovic N, et al. Role of proximal promoter elements in regulation of renin gene transcription. J. Biol. Chem. 271: 22499-22505, 1996. PubMed: 8798416

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