DLD-1 人結(jié)直腸腺癌上皮細(xì)胞
ATCC® Number: CCL-221?
Designations: DLD-1
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
Cell Type: epithelial
Cellular Products: carcinoembryonic antigen (CEA) 0.5 ng/10 exp6 cells/10 days; colon antigen 3; keratin
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Applications: transfection host (Roche FuGENE® Transfection Reagents)
Tumorigenic: Yes
Reverse Transcript: negative
Oncogene: myc +; myb + ; ras +; fos +; sis +; p53 +; abl -; ros -; src -
Antigen Expression: blood type O
DNA Profile (STR): Amelogenin: X,Y
CSF1PO: 11,12
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 pseudodiploid human cell line with the modal chromosome number of 46, occurring in 86% of cells. The rate of polyploidy was high at 17.1%. The karyotype of the line was 46,XY,-2,+dir dup(2)(p13-p23). The Y chromosome was slightly longer than N22 and had a large segment of heterochromatic, fluorescent distal q arms.
Isoenzymes: ES-D, 1-2
G6PD, B
PEP-D, 1
PGD, A
PGM1, 1
PGM3, 1
Age:
Gender: male
Comments: DLD-1 is one of two colorectal adenocarcinoma cell lines which were isolated by D.L. Dexter and associates during a period from 1977-1979 [23033]
DNA fingerprinting and cytogenetic analyses performed at ATCC and elsewhere show the line is similar to HCT-15 (CCL-225) and suggest the two are of different clonal origin from the same individual. [22417]
Their genetic origin has been confirmed by DNA fingerprinting; however, cytogenetic analysis has shown that they lack concurrent marker chromosomes or concurrent numerical changes.
The cells are negative for CSAp (CSAp-).
DLD-1 cells are positive for p53 antigen expression (the p53 antigen produced has a C -> T mutation resulting in Ser -> Phe at position 241).
The cells are positive for keratin by immunoperoxidase staining.
The line is positive for expression of c-myc, K-ras, H-ras, N-ras, myb, sis and fos oncogenes.
N-myc oncogene expression was not detected.
Tumor specific nuclear matrix proteins CC-2, CC-3, CC-4, CC-5 and CC-6 are expressed.
A culture of unknown passage submitted to the ATCC in 1979 was found to be contaminated with Mycoplasma hyorhinis. The cells were subsequently cured using a combination of antibiotics over a 12-week cultivation period.
Following treatment, the cells were assayed by the Hoechst stain weekly and by the standard culture test periodically. During 11 consecutive months of cultivation in the absence of antibiotics, all of these tests were negative.
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 medium, and rinse with 0.25% trypsin, 0.53 mM EDTA solution. Remove the solution and add an additional 1 to 2 ml of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37C) until the cells detach. Add fresh culture medium, aspirate and dispense into new culture flasks.
Subcultivation Ratio: A subcultivation ratio of 1:3 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 temperature
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2001
recommended serum:ATCC 30-2020
References: 22147: Chen TR, et al. Intercellular karyotypic similarity in near-diploid cell lines of human tumor origins. Cancer Genet. Cytogenet. 10: 351-362, 1983. PubMed: 6652615
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
23322: Rodrigues NR, et al. p53 mutations in colorectal cancer. Proc. Natl. Acad. Sci. USA 87: 7555-7559, 1990. PubMed: 1699228
23341: Keesee SK, et al. Nuclear matrix proteins in human colon cancer. Proc. Natl. Acad. Sci. USA 91: 1913-1916, 1994. PubMed: 8127905
32794: Kutchera W, et al. Protaglandin H synthase 2 is expressed abnormally in human colon cancer: evidence for a transcriptional effect. Proc. Natl. Acad. Sci. USA 93: 4816-4820, 1996. PubMed: 8643486
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
