HCT-15 人結(jié)直腸腺癌細(xì)胞
ATCC® Number: CCL-225?
Designations: HCT-15
Depositors: DL Dexter
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Homo sapiens (human)
Morphology: epithelial
Source: Organ: colon
Tumor Stage: Dukes' type C
Disease: colorectal adenocarcinoma
Cellular Products: carcinoembryonic antigen (CEA) 5.4 ng/10 exp6 cells/10 days; keratin
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Applications: transfection host (Roche FuGENE® Transfection Reagents)
Tumorigenic: Yes
Reverse Transcript: negative
DNA Profile (STR): Amelogenin: X,Y
CSF1PO: 12
D13S317: 8,11
D16S539: 12,13
D5S818: 13
D7S820: 10,12
THO1: 7,9.3
TPOX: 8,11
vWA: 18,19
Cytogenetic Analysis: This is a quasidiploid human cell line with the modal number 46 occurring in 76% of cells (range = 41 to 47 for 50 metaphases).The rate of polyploidy was 5.1%. The karyotype of the line 46, XY, -8,-11, -17, t(8:17)(p23:q21), inv(11)(p15.3q13.1). The Y chromosome was slightly longer than N22, and had a large segment of heterochromatic, fluorescent distal q arms.
Isoenzymes: ES-D, 2
G6PD, B
PEP-D, 1
PGD, A
PGM1, 1-2
PGM3, 1
Gender: male
Comments: Evidence from DNA fingerprinting indicates that this line and DLD-1 (ATCC CCL-221) are derived from the same individual; however, isoenzymology and cytogenetic data leave some doubt.
HCT-15 cells are CSAp negative (CSAp-).
The cells are positive for keratin by immunoperoxidase staining.
Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
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 that 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.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:10 is recommended
Medium Renewal: 2 to 3 times per week
Preservation: Freeze medium: Complete growth medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2001
recommended serum:ATCC 30-2020
References: 22417: Chen TR, et al. DLD-1 and HCT-15 cell lines derived separately from colorectal carcinomas have totally different chromosome changes but the same genetic origin. Cancer Genet. Cytogenet. 81: 103-108, 1995. PubMed: 7621404
22861: Trainer DL, et al. Biological characterization and oncogene expression in human colorectal carcinoma cell lines. Int. J. Cancer 41: 287-296, 1988. PubMed: 3338874
23033: Dexter DL, et al. N,N-dimethylformamide-induced alteration of cell culture characteristics and loss of tumorigenicity in cultured human colon carcinoma cells. Cancer Res. 39: 1020-1025, 1979. PubMed: 427742
32266: Bender CM, et al. Inhibition of DNA methylation by 5-Aza-2'-deoxycytidine suppresses the growth of human tumor cell lines. Cancer Res. 58: 95-101, 1998. PubMed: 9426064
46971: Vermeulen SJ, et al. Did the four human cancer cell lines DLD-1, HCT-15, HCT-8, and HRT-18 originate from one and the same patient?. Cancer Genet. Cytogenet. 107: 76-79, 1998. PubMed: 9809040
