Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimay responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Restrictions: The cells are distributed for research purposes only. The Memorial Sloan-Kettering Cancer Center releases the line subject to the following: 1.) The cells or their products must not be distributed to third parties. Commercial interests are the exclusive property of Memorial Sloan-Kettering Cancer Center. 2.) Any proposed commercial use of these cells must first be negotiated with The Director, Office of Industrial Affairs, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021; phone (212) 639-6181; (212) 717-3439.
Isolation: Isolation date: 1964
Applications: transfection host (Nucleofection technology from Lonza
Roche FuGENE® Transfection Reagents)
Receptors: human adrenergic alpha2A [23560]
urokinase receptor (u-PAR)
vitamin D (moderate expression)
urokinase receptor (u-PAR); vitamin D (moderate expression)
Virus Susceptibility: human immunodeficiency virus (HIV, LAV)
Cytogenetic Analysis: modal number = 71; range = 68 to 72.
The stemline chromosome number is hypertriploid with the 2S component occurring at 2.4%. Seventeen marker chromosomes are found in most metaphases, generally in single copy per chromosome. The marker designations are: M1p-(=t(3p-;?) with a deleted short arm), t(7q;?), t(10q;?), i(13q), 19q+a; M6, ?t(8q;9q-), ?Xp, M9, 6q+, t(13;?)a, t(13;?)b, 19q+b, M14, M15, 15p+, and Xq-. Chromosome 13 is nullisomic and chromosomes 8 and 14 are generally monosomic. No Y chromosome was detected by QM band analysis.
Isoenzymes: AK-1, 1
ES-D, 1
G6PD, B
GLO-I, 1-2
Me-2, 1
PGM1, 1-2
PGM3, 1-2
Age: 44 years adult
Gender: female
Ethnicity: Caucasian
Comments: 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.
Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated McCoy's 5a Medium Modified, Catalog No. 30-2007. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Subculturing: Protocol:
Remove and discard culture medium.
Briefly rinse the cell layer with 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37C to facilitate dispersal.
Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
Add appropriate aliquots of the cell suspension to new culture vessels.
Incubate cultures at 37C.
Subc*tion Ratio: A subc*tion ratio of 1:3 to 1:8 is recommended
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