Cellular Products: carcinoembryonic antigen (CEA) (The production of CEA in the ATCC seed stock was 1944 ng per 10(6) cells in 10 days.)
interleukin 10 (IL-10) [22511]
interleukin 6 (IL-6)
mucin
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Antigen Expression: serologically defined colon cancer antigen 3; Homo sapiens, expressed
HLA A2, B13, B50; Blood type O
Cytogenetic Analysis: 45,X; one X chromosome missing; no other chromosomal aberrations
Isoenzymes: ADA, 1
ES-D, 1
G6PD, B
PEP-D, 1
PGD, A
PGM1, 1
PGM3, 2
Age: 58 years
Gender: female
Ethnicity: Caucasian
Comments: LS 174T is a variant of LS 180 (ATCC CL-187) that has been maintained by using trypsin in the subculture protocol. It is more easily subc*ted than that parent line and, like LS 180, it is reported to produce large amounts of carcinoembryonic antigen (CEA).
Electron microscopic studies revealed abundant microvilli and intracytoplasmic mucin vacuoles [Pubmed ID: 1262041].
They are negative for p53 antigen expression, but positive for mRNA expression.
LS 174T cells stain positively for cytokeratins.
The line is positive for expression of c-myc, N-myc, H-ras, N-ras.
Myb, and fos oncogenes.
K-ras and sis oncogene expression were not detected.
Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
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.
Subc*tion Ratio: A subc*tion ratio of 1:2 to 1:4 is recommended
Medium Renewal: 2 to 3 times per week
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2003
recommended serum:ATCC 30-2020
derived from same individual:ATCC CL-187
References: 2148: Tom BH, et al. Human colonic adenocarcinoma cells. I. Establishment and description of a new line. In Vitro 12: 180-191, 1976. PubMed: 1262041
3524: Tom BH, et al. Process of producing carcinoembryonic antigen. US Patent 4,228,236 dated Oct 14 1980
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
22511: Gastl GA, et al. Interleukin-10 production by human carcinoma cell lines and its relationship to interleukin-6 expression. Int. J. Cancer 55: 96-101, 1993. PubMed: 8344757
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
32265: Tsao H, et al. Novel mutations in the p16/CDKN2A binding region of the Cyclin-dependent Kinase-4 gene. Cancer Res. 58: 109-113, 1998. PubMed: 9426066
32377: Shibata D, et al. Genomic instability in repeated sequences is an early somatic event in colorectal tumorigenesis that persists after transformation. Nat. Genet. 6: 273-281, 1994. PubMed: 8012390
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
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