DLD-1 ATCC ® CCL-221™ Homo sapiens colon Dukes' type C,colo

Permits	These permits may be required for shipping this product: Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
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 temperature

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
Oncogene	myc +; myb + ; ras +; fos +; sis +; p53 +; abl -; ros -; src -
Genes Expressed	carcinoembryonic antigen (CEA) 0.5 ng/10 exp6 cells/10 days; colon antigen 3; keratin,myc +; myb + ; ras +; fos +; sis +; p53 +; abl -; ros -; src -,blood type O,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. The cells are weakly positive for keratins and vimentin.
Tumorigenic	Yes
Comments	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. 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.

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	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 37°C) 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
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 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. [1] Chen et al. 1995 [2] Vermeulen et al., 1998 Tumors developed within 21 days at 100% frequency (5/5) in nude mice inoculated subcutaneously with 10(7) cells

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

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. [1] Chen et al. 1995 [2] Vermeulen et al., 1998

Tumors developed within 21 days at 100% frequency (5/5) in nude mice inoculated subcutaneously with 10(7) cells

Permits

These permits may be required for shipping this product:

Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.

Basic Documentation

Product Sheet

Certificate of Analysis

MSDS