2V6.11 cell line

male

This line was derived in 2001 from the human embryonic kidney line, HEK-293. HEK-293 cells were obtained by transfecting the plasmid pVgRXR containing a Zeocin?resistance selectable marker (293VgRXR cells) followed by transfecting the plasmid pEKORF6 which contians Ad5 E4 ORF 6 that encodes E4 34K. The plasmid pEKORF6 is derived from a hygromycin resistance gene containing plasmid pIndHydro, with vectors containing SV40 viral DNA. The 2V6.11 cell line was initially cloned with a cloning cylinder selected in hygromycin containing medium. The 2V6.11 cells inducibly express the human adenovirus E4, (a 34kDa protein that can be detected by western blot). The 2V6.11 can be used in the study of the adenoviral E4 oncoprotein.

2

Freeze medium: 60% Basal medium+30% FBS+10% DMSO Storage temperature: Liquid nitrogen vapor phase

MEM(PM150410)+10% FBS+1% P/S(PB180120)

Adherent

Atmosphere: Air, 95%; CO2, 5% Temperature: 37°C

Epithelial

Remove and discard culture medium. Briefly rinse the cell layer with DPBS solution to remove all traces of serum that contains trypsin inhibitor. Add 1.0 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 2 to 3 minutes). Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 4.0 to 6.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels.

Kidney;