Product Description
A cell line established from a lymph node metastasis taken from the same patient one year later is available.The line is negative for CSAp(CSAp-) and colon antigen 3.The cells are positive for keratin by immunoperoxidase staining.There is a G-】 A mutation in codon 273 of the p53 gene resulting in an Arg-】 His substitution and a C-】 T mutation in codon 309 resulting in a Pro-】 Ser substitution.The cells express elevated levels of p53 protein.The line is positive for expression of c-myc, K-ras, H-ras, N-ras, myb, sis and fos oncogenes.N-myc oncogene expression was not detected.Matrilysin, a metalloproteinase associated with tumor invasiveness, is not expressed.The cells have been reported to produce GM-CSF.
Information
ApplicationThis cell line is a suitable transfection host. This line has a mutation in codon 12 of the ras proto-oncogene, and can be used as a positive control for PCR assays of mutation in this codon.
TissueHuman colon
Morphologyepithelial
Growth Propertiesadherent
EffectsYes, in nude mice.
Specifications
Complete Growth MediumDMEM (PM150210)+10% FBS (164210-500)+1% P/S (PB180120)
Subcultivation Ratio1:2-1:4
Medium Renewalevery 2 to 3 days
CryopreservationFreeze medium: 60% Basal medium＋30% FBS＋10% DMSO
Storage temperature: Liquid nitrogen vapor phase
Culture ConditionsAtmosphere: Air, 95%; CO2, 5%
Temperature: 37℃
TumorigenicYes, Tumors developed within 21 days at 100% frequency(5/5) in nude mice inoculated subcutaneously with 10^7 cells.
Receptor Expressionepidermal growth factor(EGF)
Gene Expressioncarcinoembryonic antigen(CEA) 0.7ng/10^6 cells/10 days; keratin; transforming growth factor beta, myc＋; myb＋; ras＋; fos＋; sis＋; p53＋; abl -; ros -; src -, HLA A2, B8, B17; Blood Type A; Rh＋, The cells are positive for keratin by immunoperoxidase staining.
DurationThis cell line is a suitable transfection host. This line has a mutation in codon 12 of the ras proto-oncogene, and can be used as a positive control for PCR assays of mutation in this codon.
Misc Information
SubculturingRemove 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.