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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). 

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

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.

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.

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.

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