• 名称: Caco-2(人结肠癌肿瘤细胞)
  • 货号: CBP60025
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CBP60025

 

 

I. General information

Synonyms:

CaCo-2; CACO-2; Caco 2; CACO2; CaCo2; CaCO2; Caco2; Caco-2/ATCC

Background:

Established from the primary colon tumor (adenocarcinoma) of a 72-year-old Caucasian man in 1974. Forms moderately well differentiated adenocarcinomas consistent with colonic primary grade II, in nude mice.

Species:

Homo sapiens, human

Tissue:

Colon

Disease:

Colorectal adenocarcinoma

Gender:

Male, 72 years adult, Caucasian

Morphology:

Epithelial adherent cells; after splitting cells start to grow in colonies

Growth Mode:

Adherent

Doubling Time:

65~80hrs

DNA Profile:

Amelogenin: X
CSF1PO: 11
D13S317: 11,13,14
D16S539: 12,13
D5S818: 12,13
D7S820: 11,12/12, 12
THO1: 6
TPOX: 9,11

vWA: 16,18
Cobioer’s Cell Line Authentication Service

Culture Medium:

MEM+20%FBS+1% Non Essential Amino Acids (NEAA) + 1mM Sodium Pyruvate (NaP)
We strongly suggest to purchase the complete medium from Cobioer.

Cryopreservation medium:

90% Complete Medium+10%DMSO

Photo:

 

Receptor Expression:

This cell line expressed heat stable enterotoxin (Sta, E. coli) and epidermal growth factor (EGF).

Genes Expressed:

Keratin,retinoic acid binding protein 1,retinol binding protein 2

Cellular Products:

keratin
retinoic acid binding protein 1

retinol binding protein 2

Tumor Formation:

Yes, in nude mice; forms moderately well differentiated adenocarcinoma consistent with colonic primary (grade II)

Immunology:

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

Virus Susceptibility:

Human immunodeficiency virus 1

Comments:

Upon reaching confluence, the cells express characteristics of enterocytic differentiation [PubMed: 1939345]. Caco-2 cells express retinoic acid binding protein I and retinol binding protein II [PubMed: 9040537]

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 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.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 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

SMAD4

Homozygous

c.1051G>C

p.D351H

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=15868485; DOI=10.1007/s10565-005-0085-6; Sambuy Y., De Angelis I., Ranaldi G., Scarino M.L., Stammati A., Zucco F.; "The Caco-2 cell line as a model of the intestinal barrier: influence of cell and culture-related factors on Caco-2 cell functional characteristics."; Cell Biol. Toxicol. 21:1-26(2005).

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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