CCL-247 HCT 116 人結(jié)腸癌細(xì)胞
ATCC® Number: CCL-247?
Designations: HCT 116
Depositors: MG Brattain
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Homo sapiens (human)
Morphology: epithelial
Source: Organ: colon
Disease: colorectal carcinoma
Cellular Products: carcinoembryonic antigen (CEA) 1 ng per 10 exp6 cells per 10 days; keratin
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Applications: transfection host (Nucleofection technology from Lonza
Roche FuGENE® Transfection Reagents)
Tumorigenic: Yes
Reverse Transcript: negative
DNA Profile (STR): Amelogenin: X,Y
CSF1PO: 7,10
D13S317: 10,12
D16S539: 11,13
D5S818: 10,11
D7S820: 11,12
THO1: 8,9
TPOX: 8,9
vWA: 17,22
Cytogenetic Analysis: The stemline chromosome number is near diploid with the modal number at 45 (62%) and polyploids occurring at 6.8%. The markers 10q+ and t(?8p;18q) are present in all metaphases and t(9q;?16p-), in 80% of the cells karyotyped. N16 is monosomic in the presence of, but disomic in the absence of t(9q;?16p-). N10 and N18 are monosomic and other chromosomes from those mentioned above are disomic. Q-band observations revealed the presence of the Y chromosome, but not in all cells (50% of cells lacked the Y in G-band karyotypes).
Isoenzymes: AK-1, 1
ES-D, 1-2
G6PD, B
GLO-I, 1
PGM1, 1
PGM3, 1
Gender: male
Comments: The cells are positive for keratin by immunoperoxidase staining.
HCT 116 cells are positive for transforming growth factor beta 1 (TGF beta 1) and beta 2 (TGF beta 2) expression.
This line has a mutation in codon 13 of the ras protooncogene, and can be used as a positive control for PCR assays of mutation in this codon.
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
Growth Conditions: Growth and plating efficiency are enhanced by using a feeder layer of murine fibroblasts.
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 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.
