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TCMK-1小鼠肾上皮细胞株mouse kidney epithelial Cell Line-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心

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  • 货  号:TCMK-1小鼠肾上皮细胞株mouse kidney epithelial Cell Line
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TCMK-1 mouse kidney epithelial Cell Line

Cat No.: NTCC594182

BioVector NTCC Inc.



Organism



Mus
   musculus
, mouse



Tissue



kidney



Cell Type



SV40
   transformed



Product Format



frozen



Morphology



epithelial



Culture
   Properties



adherent



Biosafety
   Level



2 [Cells
   contain polyomavirus DNA sequences]



Disease



normal



Age



weanling



Strain



C3H/Mai



Storage
   Conditions



liquid
   nitrogen vapor phase



Karyotype



modal number =
   78; range = 64 to 157.

   All cells showed 2-6 acrocentric chromosomes with secondary constrictions and
   1-4 biarmed chromosomes per cells. Sixty percent of the cells had one or more
   minute chromosomes.



Derivation



The cells are SV40
   transformed and are positive for SV40 T-antigen.



Antigen
   Expression



H-2k



Tumorigenic



No



Effects



No, in
   immunosuppressed mice


Yes, in
   semisolid medium



Virus
   Susceptibility



Vesicular
   stomatitis, Glasgow (Indiana)

   Human poliovirus 1



Virus
   Resistance



poliovirus 1



Comments



About 2% of
   the cells are positive for SV40 viral antigen, but virus is not recoverable.


Tested and
   found negative for ectromelia virus (mousepox).



Complete
   Growth Medium



The base
   medium for this cell line is NTCC-formulated Eagle's Minimum Essential
   Medium, Catalog No. 30-2003. 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- 0.53 mM 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: A subcultivation ratio of 1:2 to 1:8 is recommended


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



Freeze medium: Complete
   growth medium 95%; DMSO, 5%


Storage
   temperature: liquid nitrogen vapor phase



Culture
   Conditions



Temperature: 37°C



References



Black PH, Rowe
   WP. SV-40 induced proliferation of tissue culture cells of rabbit, mouse, and
   porcine origin. Proc. Soc. Exp. Biol. Med. 114: 721-727, 1963. PubMed: 14120333




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