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M-1
M-1
規(guī)格:
價格:
編號:B165033
品牌:Mingzhoubio

標準菌株
定量菌液
DNA
RNA

規(guī)格:
凍干粉
斜面
甘油
平板


產(chǎn)品名稱
M-1
商品貨號
B165033
Organism
Mus musculus, transgenic for SV40 early region, mouse, transgenic for SV40 early region
Tissue
kidney, cortex, collecting duct
培養(yǎng)基
Cell Type
Epithelial
Product Format
frozen
Morphology
epithelial
Culture Properties
adherent
Biosafety Level
2 [Cells contain SV-40 viral DNA 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.

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
Deposited As
mouse, transgenic for SV40 early region
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.

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