2v6.11 Cell Line

Brief Introduction
Product List

General Information
Organism	Homo sapiens, human
Cell Line Description	2V6.11 is an engineered, ecdysone-inducible human embryonic kidney cell line. Derived from the parental HEK-293 (293) cell line, it was constructed in 2001 (and fully characterized by E.S. Mohammadi and colleagues in 2004). Its construction involved first transfecting HEK-293 cells with the plasmid pVgRXR—which carries a Zeocin resistance marker—to generate the intermediate cell line 293pVgRXR, followed by transfection with the plasmid pEKORF6; the latter contains the Adenovirus type 5 (Ad5) E4orf6 gene (encoding the E4 34K oncoprotein) under the control of an ecdysone-inducible promoter. 2V6.11 cells grow as an adherent epithelial monolayer and are recognized as a highly specific model for studying adenovirus pathology and DNA double-strand break (DSB) repair. Induced expression of the E4 34K protein disrupts the cellular non-homologous end joining (NHEJ) process and leads to the degradation of key host DNA repair proteins. Furthermore, this cell line serves as a crucial host system for the construction and propagation of complex adenoviral vectors, particularly human adenovirus type 40 (Ad40).
Tissue	Kidney
Cell Type	Embryonic Epithelial cell
Disease	Normal / Non-malignant tissue origin
Morphology	Epithelial-like
Gender	Female
Age	Fetal
Product Format	Frozen
Growth Mode	Adherent Monolayer
Biosafety Level	2 (BSL-2 is required globally because it is a HEK-293 derivative harboring integrated adenoviral and SV40 sequence constructs)
Applications	1. Inhibition of DNA double-strand break (DSB) repair and direct in vitro modeling of the non-homologous end joining (NHEJ) pathway; 2. Biochemical characterization and analysis of the degradation network of the adenoviral E4 34K oncoprotein; 3. High-efficiency vector packaging and amplification of fastidious human adenovirus strains (e.g., adenovirus type 40); 4. Host systems for evaluating ecdysone-inducible/regulated expression systems; 5. Investigation of proteasome-mediated degradation pathways targeting host proteins such as Mre11 and DNA ligase IV.
Shipped In	Dry ice
Storage Temperature	−196°C
Characteristics
Tumorigenic	No, derived from the non-tumorigenic parent HEK-293 cell line.
Plasmid Constructs	1. pVgRXR: Confers Zeocin-resistance and delivers the heterodimeric ecdysone receptor. 2. pEKORF6: Derived from pIndHydro; contains a hygromycin resistance gene and SV40 viral DNA elements.
Inducible Expression	Addition of an inducer (such as muristerone A or ponasterone A) triggers robust transcription of the E4 34K (34 kDa) protein. This expression leads to the rapid, proteasome-mediated degradation of DNA repair proteins, including Mre11 and DNA ligase IV.
Growth Kinetics	Highly proliferative adherent expansion; typical population doubling time spans approximately 24 to 30 hours under standard conditions.
Mycoplasma Test	Negative
Culture Conditions and Handling
Subculturing	1. Remove and discard the spent complete culture medium from the culture vessel. 2. Briefly rinse the cell monolayer with sterile Ca2+/Mg2+-free PBS (or DPBS) to remove any residual serum containing active trypsin inhibitors. 3. Add 1.0 to 2.0 mL of 0.25% trypsin-0.53 mM EDTA solution (or Accutase) to completely cover the cell layer. 4. Incubate at room temperature or 37°C for 2 to 4 minutes; monitor under an inverted microscope until the cells become rounded, loosen, and detach. 5. Add an equal volume of pre-warmed complete culture medium to rapidly neutralize the enzymatic reaction. 6. Centrifuge the cell suspension at ~300 × g for 3 minutes, discard the supernatant, resuspend the cell pellet uniformly in fresh complete culture medium, and distribute into new vessels.
Thawing Protocol Note	During the early recovery phase following thawing, 2V6.11 cells are highly sensitive to sudden changes in culture medium alkalinity. Before adding the thawed cell suspension, the culture vessel containing fresh complete growth medium should be placed in a 5% CO2 incubator for at least 15 minutes to allow the medium to reach and equilibrate within the normal physiological pH range (7.0 to 7.6).
Medium Renewal	2 to 3 times per week
Subcultivation Ratio	Split optimal, confluent cultures at a standard ratio of 1:3 to 1:8.
Culture Conditions	Atmosphere: Air, 95%; CO2, 5%; Temperature: 37°C
Cryopreservation	90% Complete culture growth medium + 10% DMSO (or 60% Basal medium + 30% FBS + 10% DMSO)

Cat.No.	Product Name	Price
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CSC-RR00893	GFP Reporter Cell Line - 2v6.11	Inquiry
CSC-RR00942	Luciferase Reporter Cell Line - 2v6.11	Inquiry

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* For research use only. Not intended for any clinical use.

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