Human Cell Lines-Cell Lines-InCellGene LLC.-CellLines,Biomedicine reasearch Product,Cell Biology,Cell Culture,293T Cell,Media

Permits and Restrictions	View Restrictions
Organism	Homo sapiens, human
Tissue	colon
Product Format	frozen
Morphology	epithelial
Culture Properties	adherent
Biosafety Level	1 Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Disease	colorectal adenocarcinoma
Age	44 years adult
Gender	female
Ethnicity	Caucasian
Applications	This cell line is a suitable transfection host.
Storage Conditions	liquid nitrogen vapor temperature

Karyotype	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.

Derivation	The HT-29 line was isolated from a primary tumor in 1964 by J. Fogh using the explant culture method.
Clinical Data	44 years adult Caucasian female
Antigen Expression	Blood Type A; Rh+; HLA A1, A3, B12, B17, Cw5
Receptor Expression	human adrenergic alpha2A Ref, urokinase receptor (u-PAR), vitamin D (moderate expression), urokinase receptor (u-PAR); vitamin D (moderate expression), human adrenergic alpha2A Ref
Oncogene	myc +; ras +; myb +; fos +; sis +; p53 +; abl -; ros -; src -
Genes Expressed	secretory component of IgA; carcinoembryonic antigen (CEA); transforming growth factor beta binding protein; mucin,myc +; ras +; myb +; fos +; sis +; p53 +; abl -; ros -; src -,Blood Type A; Rh+; HLA A1, A3, B12, B17, Cw5,HT-29 cells are negative for CD4, but there is cell surface expression of galactose ceramide (a possible alternative receptor for HIV).
Cellular Products	secretory component of IgA; carcinoembryonic antigen (CEA); transforming growth factor beta binding protein; mucin
Tumorigenic	Yes
Effects	Yes, in nude mice; forms well differentiated adenocarcinoma consistent with colonic primary (grade I); tumors also form in steroid treated hamsters
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.

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%.
Subculturing	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). Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product. 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 37°C 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 37°C. Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended Medium Renewal: 2 to 3 times per week
Cryopreservation	Freeze medium: Complete growth medium, 95%; DMSO, 5% Storage temperature: liquid nitrogen vapor temperature
Culture Conditions	Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C