The cells contain a stably integrated human papilloma virus(HPV) E6 oncogene under control of the cytomegalovirus(CMV) promoter. The HPV E6 oncogene causes a decrease in normal p53 levels and functions.The RKO-E6 cell line lacks appreciable functional p53.Disruption of normal p53 function in human colon carcinoma RKO cells with the human papillomavirus E6 oncoprotein results in reduced repair of u.v.-induced DNA damage and also loss of induced repair following cellular u.v.-irradiation.RKO cells contain wild-type p53 but lack endogenous human thyroid receptor nuclear receptor(h-TRbeta1). The level of p53 protein is higher in RKO cells than in RKO-E6 cells.The RKO-E6 cell line can be used together with its parental cell line, RKO to investigate the effects of p53 loss on cellular parameters such as p53 mediated transcription and apoptosis.
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.
Freeze medium: 60% Basal medium+30% FBS+10% DMSO Storage temperature: Liquid nitrogen vapor phase
For cells, cell lines and tissues in culture till half confluency.
MEM (PM150410)+10% FBS (164210-500)+1% P/S (PB180120)
Atmosphere: Air, 95%; CO2, 5% Temperature: 37℃
Liquid nitrogen -196°C
Every 2 to 3 days
Epithelial
Adherent
Dry ice
3-5 days
1:2-1:4
RKO-E6
Colon
NA
NA
NA
NA
NA
NA
NA
NA
NA
2