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

Organism	Homo sapiens, human
Tissue	colon; derived from metastatic site: ascites
Product Format	frozen
Morphology	epithelial
Culture Properties	mixed, adherent and suspension
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 D, colorectal adenocarcinoma
Age	70 years
Gender	male
Ethnicity	Caucasian
Applications	This cell line is a suitable transfection host.
Storage Conditions	liquid nitrogen vapor phase

Karyotype	The stemline chromosome number is hypertriploid and 10-11 marker chromosomes (M1, M2, M4, M5, M6 and 5 others) were common in monosomic, disomic and trisomic condition although most of these markers were monosomic.
Derivation	This line was isolated in 1975 by T.U. Semple, et al. from ascitic fluid of a 70-year-old Caucasian male with carcinoma of the colon. The patient had been treated with 5-fluorouracil for 4-6 weeks before removal of the fluid specimen.
Clinical Data	70 years Caucasian male The patient had been treated with 5-fluorouracil for 4-6 weeks before removal of the fluid specimen.
Genes Expressed	carcinoembryonic antigen (CEA) 1.5 to 4.1 ng/10 exp6 cells/10 days; keratin; interleukin 10 (IL-10, interleukin-10),The cells are positive for keratin by immunoperoxidase staining.
Cellular Products	carcinoembryonic antigen (CEA) 1.5 to 4.1 ng/10 exp6 cells/10 days; keratin; interleukin 10 (IL-10, interleukin-10)
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	The line was derived from tissue from the same patient as COLO 201 (ATCC CCL-224). The cells are CSAp negative (CSAp-). The cells are positive for keratin by immunoperoxidase staining. COLO 205 cells express a 36000 dalton cell surface glycoprotein related to the GA733-2 tumor associated antigen.

Complete Growth Medium	The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, ATCC 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (ATCC 30-2020) to a final concentration of 10%.
Subculturing	Cells grow loosely attached and in suspension. Shake flask, retain the floating cells by transfering them into a centrifuge tube. Cells that remain attached may be removed using a standard trypsinization protocol (see below) and combined with the retained floating cells. Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum which, 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 10 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 the cell suspension to the centrifuge tube containing the floating cells. Centrifuge at 125 X g for 5 to 10 minutes. Resuspend in fresh medium and plate at the appropriate subcultivation ratio. Incubate cultures at 37°C. Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:6 is recommended Medium Renewal: Every 2 to 3 days
Cryopreservation	Freeze medium: Complete culture medium, 95%; DMSO, 5% Storage temperature: liquid nitrogen vapor phase
Culture Conditions	Temperature: 37°C