細(xì)胞類型：其他細(xì)胞類型
是否是腫瘤細(xì)胞：0
物種來源：小鼠
年限：adult
細(xì)胞形態(tài)：其他
生長狀態(tài)：貼壁生長
運輸方式：凍存運輸
數(shù)量：大量
器官來源：大動脈
品系：C57BL/6
組織來源：smooth muscle
ATCC Number：CRL-2797?
規(guī)格：0.2ml Designations: MOVAS
Depositors: ?M Husain
Biosafety Level:2
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism: Mus musculus
Morphology:spindle-shaped
Source: Organ: aorta
Strain: C57BL/6
Tissue: smooth muscle
Cell Type: immortalized with SV40 large T antigen
Cellular Products:actin; smooth muscle alpha actin
calponin 1; basic; smooth muscle
desmin
smooth muscle protein 22 (SM22 alpha)
Permits/Forms:In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Isolation: Isolation date: December, 1999
Age: adult
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.
Propagation: ATCC 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%
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Subculturing: Protocol:
- 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 gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 4 X 10(3) to 2 X 10(4) viable cells/sq. cm is recommended.
- Incubate cultures at 37?C.
Interval: Maintain cultures at a cell concentration between 1 X 10(4) and 1 X 10(5) cells/sq. cm
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended
Medium Renewal: Two to three times weekly
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Doubling Time: 15 hours
Related Products:Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
recommended serum:ATCC 30-2020
Cell culture tested DMSO:ATCC 4-X
Erythrosin B vital stain solution:ATCC 30-2404
References: 47767: 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
88855: 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
