• 名称: SW480(人结肠癌细胞)
  • 货号: CBP60019
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CBP60019  

Synonyms:

SW-480; SW 480; SW480E

Background:

Established from the tumor of a 50-year-old Caucasian man with colon adenocarcinoma (grade 4, Duke class B)

Species:

Homo sapiens, human

Tissue:

Colon

Disease:

Dukes' type B, colorectal adenocarcinoma

Gender:

Male, 50 years adult, Caucasian

Morphology:

Endothelial-like adherent cells, mostly bipolar with microvilli growing in monolayers

Growth Mode:

Adherent

Doubling Time:

25~30hrs

DNA Profile:

Amelogenin: X
CSF1PO: 13,14
D13S317: 12
D16S539: 13
D5S818: 13
D7S820: 8
THO1: 8
TPOX: 11
vWA: 16
Cobioer’s Cell Line Authentication Service

Culture Medium:

DMEM+10%FBS
We strongly suggest to purchase the complete medium from Cobioer.

Cryopreservation medium:

90% Complete Medium+10%DMSO

Photo:

 

Receptor Expression:

Epidermal growth factor (EGF)

Oncogene:

Myc +; myb + ; ras +; fos +; sis +; p53 +; abl -; ros -; src -

Genes Expressed:

carcinoembryonic antigen (CEA) 0.7 ng/10 exp6 cells/10 days; keratin; transforming growth factor beta,myc +; myb + ; ras +; fos +; sis +; p53 +; abl -; ros -; src -,HLA A2, B8, B17; blood type A; Rh+,The cells are positive for keratin by immunoperoxidase staining.,The line is positive for expression of c-myc, K-ras, H-ras, N-ras, myb, sis and fos oncogenes.

Cellular Products:

carcinoembryonic antigen (CEA) 0.7 ng/10 exp6 cells/10 days; keratin; transforming growth factor beta

Tumor Formation:

Yes. Tumors developed within 21 days at 100% frequency (5/5) in nude mice inoculated subcutaneously with 10(7) cells.

Immunology:

Cytokeratin +, cytokeratin-7 -, cytokeratin-8 +, cytokeratin-17 -, cytokeratin-18 +, cytokeratin-19 +, desmin -, endothel -, EpCAM +, GFAP -, neurofilament -, vimentin +

Virus Susceptibility:

Human immunodeficiency virus 1 , Human immunodeficiency virus 1

Comments:

A cell line established from a lymph node metastasis taken from the same patient one year later is available (see ATCC CCL-227).
The line is negative for CSAp (CSAp-) and colon antigen 3.
The cells are positive for keratin by immunoperoxidase staining.
There is a G -> A mutation in codon 273 of the p53 gene resulting in an Arg -> His substitution and a C -> T mutation in codon 309 resulting in a Pro -> Ser substitution.
The cells express elevated levels of p53 protein.
The line is positive for expression of c-myc, K-ras, H-ras, N-ras, myb, sis and fos oncogenes.
N-myc oncogene expression was not detected.
Matrilysin, a metalloproteinase associated with tumor invasiveness, is not expressed.
The cells have been reported to produce GM-CSF

For more information, please contact Cobioer (4008-750-250).

 

II. Handling Procedure for Flask Cultures

The flask was seeded with cells grown and completely filled with medium at Cobioer 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 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 without CO2 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 without CO2.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended
Medium Renewal: 1 to 2 times per week

 

IV. Cryopreservation Procedure

Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
1.Remove and discard culture medium.
2.Briefly rinse the cell layer with 0.25% (w/v) Trypsin0.53mM EDTA solution to remove all traces of serum which 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 (usually within 1 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.
4.Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5.To remove trypsin-EDTA solution, transfer cell suspension to a centrifuge tube and spin at approximately 1000rpmfor 5 to10 minutes.
6.Discard supernatant and resuspend cells in cryopreservation medium.
7.Transfer the cells into Cryogenic Vials, 1ml/vial.
8.Frozen the cells in cryogenic container (Nalgene #5100-0001).

 

V. Database

Mutation:

Mutation Gene

Zygosity

Gene Sequence

Protein Sequence

ACSL6

Heterozygous

c.556A>C

p.I186L

APC

Homozygous

c.4012C>T

p.Q1338*

ATP1A1

Heterozygous

c.1194T>C

p.H398H

CEBPA

Heterozygous

c.976A>T

p.K326*

For more information, Please visit
http://www.cobioer.com/sjk/&pmcId=77.html

Expression:

NA
For more information, Please visit
http://www.cobioer.com/sjk/&pmcId=77.html

Comments:

For more information, please contact Cobioer (4008-750-250).

 

VI. References

1.PubMed=19927377; DOI=10.1002/gcc.20730; Knutsen T., Padilla-Nash H.M., Wangsa D., Barenboim-Stapleton L., Camps J., McNeil N., Difilippantonio M.J., Ried T.; "Definitive molecular cytogenetic characterization of 15 colorectal cancer cell lines."; Genes Chromosomes Cancer 49:204-223(2010).

2.PubMed=20606684; DOI=10.1038/sj.bjc.6605780; Bracht K., Nicholls A.M., Liu Y., Bodmer W.F.; "5-Fluorouracil response in a large panel of colorectal cancer cell lines is associated with mismatch repair deficiency."; Br. J. Cancer 103:340-346(2010).

3.PubMed=24042735; DOI=10.1038/oncsis.2013.35; Ahmed D., Eide P.W., Eilertsen I.A., Danielsen S.A., Eknaes M., Hektoen M., Lind G.E., Lothe R.A.; "Epigenetic and genetic features of 24 colon cancer cell lines."; Oncogenesis 2:E71-E71(2013).

4.PubMed=24755471; DOI=10.1158/0008-5472.CAN-14-0013; Mouradov D., Sloggett C., Jorissen R.N., Love C.G., Li S., Burgess A.W., Arango D., Strausberg R.L., Buchanan D., Wormald S., O'Connor L., Wilding J.L., Bicknell D., Tomlinson I.P.M., Bodmer W.F., Mariadason J.M., Sieber O.M.; "Colorectal cancer cell lines are representative models of the main molecular subtypes of primary cancer."; Cancer Res. 74:3238-3247(2014).
5.PubMed=25485619; DOI=10.1038/nbt.3080; Klijn C., Durinck S., Stawiski E.W., Haverty P.M., Jiang Z., Liu H., Degenhardt J., Mayba O., Gnad F., Liu J., Pau G., Reeder J., Cao Y., Mukhyala K., Selvaraj S.K., Yu M., Zynda G.J., Brauer M.J., Wu T.D., Gentleman R.C., Manning G., Yauch R.L., Bourgon R., Stokoe D., Modrusan Z., Neve R.M., de Sauvage F.J., Settleman J., Seshagiri S., Zhang Z.; "A comprehensive transcriptional portrait of human cancer cell lines."; Nat. Biotechnol. 33:306-312(2015).
6.PubMed=26537799; DOI=10.1074/mcp.M115.051235; Holst S., Deuss A.J.M., van Pelt G.W., van Vliet S.J., Garcia-Vallejo J.J., Koeleman C.A.M., Deelder A.M., Mesker W.E., Tollenaar R.A., Rombouts Y., Wuhrer M.; "N-glycosylation profiling of colorectal cancer cell lines reveals association of fucosylation with differentiation and caudal type homebox 1 (CDX1)/villin mRNA expression."; Mol. Cell. Proteomics 15:124-140(2016).

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