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