DLD-1 (ATCC® CCL-221)

Organism: Homo sapiens, human  /  Cell Type: epithelial  /  Tissue: colon  /  Disease: Dukes' type C, colorectal adenocarcinoma

Organism Homo sapiens, human
Tissue colon
Cell Type epithelial
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease Dukes' type C, colorectal adenocarcinoma
Age adult
Gender male
Applications
This cell line is a suitable transfection host.
Storage Conditions liquid nitrogen vapor phase
Karyotype 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.
Derivation
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
Clinical Data
male
Antigen Expression

Blood type O.

The cells are weakly positive for keratins and vimentin. The cells are positive for keratin by immunoperoxidase staining.

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). 

Oncogene myc +; myb + ; ras +; fos +; sis +; p53 +; abl -; ros -; src -
Genes Expressed

carcinoembryonic antigen (CEA) 0.5 ng/106 cells/10 days; colon antigen 3.

Cellular Products
carcinoembryonic antigen (CEA) 0.5 ng/10 exp6 cells/10 days; colon antigen 3; keratin
Tumorigenic Yes
Effects
Yes, in nude mice
Comments

This cell line is one of four colorectal adenocarcinoma cell lines that have been identified as being derived from a single individual. DNA profiling studies [1,2] have shown that DLD-1 (ATCC CCL-221), HCT-15 (ATCC CCL-225), HCT-8 (ATCC CCL-244) and HRT-18G (ATCC CRL-11663) share a single profile. 

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.

Their genetic origin has been confirmed by DNA fingerprinting; however, cytogenetic analysis has shown that they lack concurrent marker chromosomes or concurrent numerical changes.

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.
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%.
Subculturing

Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum, which contains trypsin inhibitor.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. 
  5. Add appropriate aliquots of the cell supension to new culture vessels.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:10 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
Temperature: 37°C
STR Profile
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
Isoenzymes
ES-D, 1-2
G6PD, B
PEP-D, 1
PGD, A
PGM1, 1
PGM3, 1
Name of Depositor DL Dexter
Passage History
A culture of unknown passage submitted to the ATCC in 1979 was found to be contaminated with Mycoplasma hyorhinis. ­
References

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

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

Trainer DL, et al. Biological characterization and oncogene expression in human colorectal carcinoma cell lines. Int. J. Cancer 41: 287-296, 1988. PubMed: 3338874

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

Rodrigues NR, et al. p53 mutations in colorectal cancer. Proc. Natl. Acad. Sci. USA 87: 7555-7559, 1990. PubMed: 1699228

Keesee SK, et al. Nuclear matrix proteins in human colon cancer. Proc. Natl. Acad. Sci. USA 91: 1913-1916, 1994. PubMed: 8127905

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

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

Basic Documentation
References

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

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

Trainer DL, et al. Biological characterization and oncogene expression in human colorectal carcinoma cell lines. Int. J. Cancer 41: 287-296, 1988. PubMed: 3338874

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

Rodrigues NR, et al. p53 mutations in colorectal cancer. Proc. Natl. Acad. Sci. USA 87: 7555-7559, 1990. PubMed: 1699228

Keesee SK, et al. Nuclear matrix proteins in human colon cancer. Proc. Natl. Acad. Sci. USA 91: 1913-1916, 1994. PubMed: 8127905

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

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