產(chǎn)品名稱 |
MOVAS |
商品貨號 |
B220580 |
Organism |
Mus musculus, mouse |
Tissue |
aorta/smooth muscle |
Cell Type |
immortalized with SV40 large T antigen |
Product Format |
frozen |
Morphology |
spindle-shaped |
Culture Properties |
adherent |
Biosafety Level |
2
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. |
Age |
adult |
Strain |
C57BL/6 |
Storage Conditions |
liquid nitrogen vapor phase |
Images |
|
Derivation |
Primary vascular aortic smooth muscle cells isolated by collagenase - elastase digestions in December 1999 were transduced with retrovirus encoding the SV40 large T antigen as well as the gene for neomycin resistance. Clones were selected in the presence of 0.4 mg/ml G418 and subcloned by limiting dilution. |
Genes Expressed |
actin; smooth muscle alpha actin,calponin 1; basic; smooth muscle,desmin,smooth muscle protein 22 (SM22 alpha) |
Cellular Products |
actin; smooth muscle alpha actin calponin 1; basic; smooth muscle desmin smooth muscle protein 22 (SM22 alpha) |
Comments |
Primary vascular aortic smooth muscle cells isolated by collagenase - elastase digestions in December 1999 were transduced with retrovirus encoding the SV40 large T antigen as well as the gene for neomycin resistance. Clones were selected in the presence of 0.4 mg/ml G418 and subcloned by limiting dilution. |
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:
0.2 mg/ml G -418
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.
- Remove and discard culture medium.
- 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.
- 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.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by pipetting gently.
- Add appropriate aliquots of cell suspension to new culture vessels. An inoculum of 4 x 103 to 2 x 104 viable cells/cm2 is recommended. Maintain cultures at a cell concentration between 1 X 104 and 1 X 105 cells/cm2
- Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended
Medium Renewal: Two to three times weekly
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture of Animal Cells: a Manual of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005 |
Cryopreservation |
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase |
Culture Conditions |
Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C |
Population Doubling Time |
15 hours |
Name of Depositor |
M Husain |
Year of Origin |
December, 1999 |
References |
Afroze T, Husain M. c-Myb-binding sites mediate G(1)/S-associated repression of the plasma membrane Ca(2+)-ATPase-1 promoter. J. Biol. Chem. 275: 9062-9069, 2000. PubMed: 10722757
Afroze T, et al. Calcineurin-independent regulation of plasma membrane Ca2+ ATPase-4 in the vascular smooth muscle cell cycle. Am. J. Physiol. Cell Physiol. : C88-C95, 2003. PubMed: 12660151
|