CRL-2784 2V6.11 JHU-67 人胚腎細(xì)胞

ATCC® Number:  CRL-2784™

Designations:  2V6.11 [JHU-67]

Depositors:   G Ketner

Biosafety Level: 2 [Cells containing Adenovirus viral DNA sequences ]

Shipped:  frozen

Medium & Serum:  See Propagation

Growth Properties: adherent

Organism: Homo sapiens (human)

Morphology: epithelial

CRL-2784 2V6.11 JHU-67 人胚腎細(xì)胞

Source: Organ: kidney

Cell Type: epithelialtransformed with adenovirus 5 DNA

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 ultimay responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.

Restrictions: Part of the Johns Hopkins Special Collection

Isolation:  Isolation date: 2001

Age:  fetus

Comments: This line was derived from the human embryonic kidney line, 293 in 2001. 293 cells were transfected with the plasmid pVgRXR bearing a Zeocin-resistance selectable marker to obtain 293VgRXR cells. This population was subsequently transfected with the plasmid pEKORF6 containing Ad5 E4 ORF 6 that encodes E4 34K. pEKORF6 is a derivative of the plasmid pIndHydro that contains a hygromycin resistance gene. The vectors contain SV40 viral DNA sequences. The 2V6.11 cell line was originally selected in hygromycin-containing medium and cloned by single-colony isolation with a cloning cylinder. The 2V6.11 cells inducibly express the human adenovirus E4, 34kDa protein (E4 ORF6). 2V6.11 cells exhibit little or no constitutive E4 biologic activity. Induction by ponasterone A, an ecdysone analog, however, results in E4 biologic activity and levels of E4 34kDa protein that are detectable by immunoblotting. These cells can be used as a tool to study the biology of the adenoviral E4 oncoprotein.

Propagation:  ATCC complete growth medium: The base medium for this cell line is ATCC-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%.

Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Temperature: 37.0°C

Growth Conditions: The depositor recommends periodic hygromycin selection.

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 5 X 10(3) to 1 X 10(4) viable cells/cm2 is recommended.

Incubate cultures at 37°C.

Interval: Maintain cultures at a cell concentration between 2 X 10(4) and 2 X 10(5) cells/cm2.

Subc*tion Ratio: A subc*tion ratio of 1:6 to 1:10 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:  23 hrs.

Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2003

recommended serum:ATCC 30-2020

source culture:ATCC JHU-67

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