>細胞CELL>biobw自建細胞系>COLO-678(人結腸癌細胞)
COLO-678(人結腸癌細胞)
產品類別:人源細胞系
生長特性:貼壁生長
細胞形態(tài):圓形或紡錘形貼壁細胞,單層生長
凍存條件:90%FBS+10%DMSO
Synonyms:
COLO-678(人結腸癌細胞)Colo-678; COLO-678; COLO678
Background:
Established from a metastatic lymph node of a 69-year-old man with colon carcinoma in 1984
Species:
Homo sapiens, human
Tissue:
Colon
Disease:
Colorectal adenocarcinoma, derived from metastatic site: Lymph node.
Gender:
Male, 69 years
Morphology:
Round or spindle-shaped adherent cells growing as monolayer
Growth Mode:
Adherent
Doubling Time:
60~70hrs
DNA Profile:
Amelogenin: X
CSF1PO: 12, 12
D13S317: 10, 13
D16S539: 9, 9
D5S818: 13, 14
D7S820: 8, 12
THO1: 9, 9.3
TPOX: 8, 8
vWA: 16, 19
Culture Medium:
RPMI-1640+10%FBS
Cryopreservation medium:
90% Complete Medium+10%DMSO
Photo:
NA
Comments:
Immunology: cytokeratin +, cytokeratin-7 -, cytokeratin-8 +, cytokeratin-17 -, cytokeratin-18 +, cytokeratin-19 +, desmin -, endothel -, EpCAM +, GFAP -, neurofilament -, vimentin –
Viruses: ELISA: reverse transcriptase negative; PCR: EBV -, HBV -, HCV -, HHV-8 -, HIV -, HTLV-I/II -, MLV -, SMRV -
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II. Handling Procedure for Flask Cultures
The flask was seeded with cells grown and completely filled with medium at to prevent loss of cells during shipping.
1.Upon receipt visually examine the culture for macroscopic evidence of any microbial contamination.
Using an inverted microscope (preferably equipped with phasecontrast optics), carefully check for any evidence of microbial contamination. Also check to determine if the majority of cells are still attached to the bottom of the flask? during shipping the cultures are sometimes handled roughly and many of the cells often detach and become suspended in the culture medium (but are still viable).
2.If the cells are still attached, aseptically remove all but 5 to 10 mL of the shipping medium. The shipping medium can be saved for reuse. Incubate the cells at 37°C in a 5% CO2 in air atmosphere until they are ready to be subcultured.
3.If the cells are not attached, aseptically remove the entire contents of the flask and centrifuge at 1000rpm for 5 to 10 minutes. Remove shipping medium and save. Resuspend the pelleted cells in 10 mL of this medium and add to 25 cm2 flask. Incubate at 37°C in a 5% CO2 in air atmosphere until cells are ready to be subcultured.
III. Subculturing Procedure
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
1.Remove and discard culture medium.
2.Briefly rinse the cell layer with 0.25% (w/v) Trypsin0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
3.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 (depend on the batch of trypsin).
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.
4.Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5.Add appropriate aliquots of the cell suspension to new culture vessels.
6.Incubate cultures at 37°C.
Note: Subculture at 85% of confluence (about every 6 to 7 days).
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:5 is recommended
Medium Renewal: 2 to 3 times per week