Human Cell Lines-Cell Lines-InCellGene LLC.-CellLines,Biomedicine reasearch Product,Cell Biology,Cell Culture,293T Cell,Media

Organism	Homo sapiens, human
Tissue	colon
Product Format	frozen
Morphology	epithelial
Culture Properties	adherent
Biosafety Level	1 Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Disease	Dukes'''''''' type B,colorectal adenocarcinoma
Age	58 years
Gender	female
Ethnicity	Caucasian
Storage Conditions	liquid nitrogen vapor phase
Disclosure	This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC.

Karyotype	45,X; one X chromosome missing; no other chromosomal aberrations

Derivation	The cell line was established from a Duke''''''''s type B adenocarcinoma of the colon. The tissue was minced and cultured without transfer for 10 months.
Clinical Data	58 years Caucasian female
Antigen Expression	serologically defined colon cancer antigen 3; Homo sapiens, expressed HLA A2, B13, B50; Blood type O
Oncogene	myc +; myb + ; ras +; fos +; p53 +; sis -; abl -; ros -; src -
Genes Expressed	carcinoembryonic antigen (CEA), interleukin 10 (IL-10), interleukin 6 (IL-6), mucin The production of CEA in the ATCC seed stock was 1944 ng per 10(6) cells in 10 days.
Cellular Products	carcinoembryonic antigen (CEA) interleukin 10 (IL-10) interleukin 6 (IL-6) mucin
Tumorigenic	Yes
Effects	Yes, in nude mice (Tumors developed within 21 days at 100% frequency (5/5) in nude mice inoculated subcutaneously with 10(7) cells)
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 subcultivated 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. Ref 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. Ref K-ras and sis oncogene expression were not detected.

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%.
Subculturing	Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. 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. Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended Medium Renewal: 2 to 3 times per week
Cryopreservation	Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Culture Conditions	Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C