HT-29 Cell Line

44 years adult

Transfection.

Ultrastructural features reported for HT-29 cells include microvilli, microfilaments, large vacuolated mitochondria with dark granules, smooth and rough endoplasmic reticulum with free ribosomes, lipid droplets, few primary and many secondary lysosomes. The cells express urokinase receptors, but do not have detectable plasminogen activator activity [PubMed ID: 8381394]. HT-29 cells are negative for CD4, but there is cell surface expression of galactose ceramide(a possible alternative receptor for HIV). The line is positive for expression of c-myc, K-ras, H-ras, N-ras, Myb, sis and fos oncogenes. The p53 antigen is overproduced, and there is a G- A mutation in codon 273 of the p53 gene resulting in an Arg- His substitution. N-myc oncogene expression was not detected.There is a G- A mutation in codon 273 of the p53 gene resulting in an Arg- His substitution.

BSL-1

Cancer cell line

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

McCoy?s 5A(PM150710)+10% FBS+1% P/S(PB180120)

~36-60 hours

Yes, in nude mice forms well differentiated adenocarcinoma consistent with colonic primary(grade I); tumors also form in steroid treated hamsters.

Adherent

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

37°C

Every 2 to 3 days

Epithelial-like

1:2-1:4

1:2 to 1:4

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.

Colon

Yes