M-1 (ATCC® CRL-2038)

Organism: Mus musculus, transgenic for SV40 early region, mouse, transgenic for SV40 early region  /  Cell Type: Epithelial  /  Tissue: kidney, cortex, collecting duct  / 

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Organism Mus musculus, transgenic for SV40 early region, mouse, transgenic for SV40 early region
Tissue kidney, cortex, collecting duct
Cell Type Epithelial
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 [Cells contain SV-40 viral DNA sequences]
Applications This cell line is a suitable transfection host
Storage Conditions liquid nitrogen vapor phase
Derivation The M-1 cell line was established from normal renal tissue taken from a mouse transgenic for the SV40 early region (tg(SV40E)Bri7).
Comments

The cells retain many characteristics of cortical collecting duct (CCD) cells including morphology and CCD antigens.

Most cell lines cloned from M-1 exhibit characteristics of either intercalated cells (ICC) or principle cells (PC) of the CCD.

5 to 10% of the cells exhibit a dual PC - ICC phenotype.

When grown on permeable supports, the cells develop a lumen negative transepithelial potential difference.

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 adjusted to contain 15 mM HEPES , 0.5 mM sodium pyruvate and 1.2 g/L sodium bicarbonate supplemented with 0.005 mM dexamethasone and 5% fetal bovine serum
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- 0.53 mM EDTA solution to remove all traces of serum that 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: A subcultivation ratio of 1:3 to 1:4 is recommended
Medium Renewal: 2 to 3 times per week

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 supplemented with 5% (v/v) DMSO
Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
Name of Depositor G Fejes-Toth
References

Fejes-Toth G, Naray-Fejes-Toth A. Differentiation of renal beta-intercalated cells to alpha-intercalated and principal cells in culture. Proc. Natl. Acad. Sci. USA 89: 5487-5491, 1992. PubMed: 1608958

Stoos BA, et al. Characterization of a mouse cortical collecting duct cell line. Kidney Int. 39: 1168-1175, 1991. PubMed: 1654478

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.

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

Fejes-Toth G, Naray-Fejes-Toth A. Differentiation of renal beta-intercalated cells to alpha-intercalated and principal cells in culture. Proc. Natl. Acad. Sci. USA 89: 5487-5491, 1992. PubMed: 1608958

Stoos BA, et al. Characterization of a mouse cortical collecting duct cell line. Kidney Int. 39: 1168-1175, 1991. PubMed: 1654478

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.