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

Permits and Restrictions	View Restrictions
Organism	Homo sapiens, human
Tissue	cecum
Product Format	frozen
Morphology	epithelial
Culture Properties	floating aggregates of round cells with some attached cells
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	colorectal adenocarcinoma
Age	55 years
Gender	male
Ethnicity	Caucasian

Karyotype	modal number = 102; range = 71 to 131; most cells have DM''s
Derivation	This line was derived from a metastasis to the abdominal wall obtained from a patient after treatment with 5-fluorouracil.
Clinical Data	55 years Caucasian male
Antigen Expression	Blood type A; Rh+; CA19-9 antigen
Genes Expressed	carcinoembryonic antigen (CEA), 1709 ng/mL per 106 cells per 10 days
Cellular Products	carcinoembryonic antigen (CEA), 1709 ng/mL per 106 cells per 10 days
Tumorigenic	Yes
Effects	Yes, in nude mice Tumors developed within 21 days at 100% frequency (5/5) in nude mice inoculated subcutaneously with 107 cells
Comments	The cells contain Dopa decarboxylase, CA19-9 antigen and CEA but do not express the TAG-72 antigen.

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	Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Remove culture medium. Keep floating cells. 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 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 and the harvested cells to new culture vessels. Incubate cultures at 37°C. Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended Medium Renewal: 3 times per week
Cryopreservation	Culture medium, 95%; DMSO, 5%

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

Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

Remove culture medium. Keep floating cells.
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 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 and the harvested cells to new culture vessels.
Incubate cultures at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended

Medium Renewal: 3 times per week

Cryopreservation

Culture medium, 95%; DMSO, 5%