• 名称: HT-29(人结肠癌细胞)
  • 货号: CBP60011
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CBP600011

 

 

I. General information

Synonyms:

HT 29; HT29

Background:

Established from the primary tumor of a 44-year-old Caucasian woman with colon adenocarcinoma in 1964; described to be heterotransplantable forming well-differentiated grade I tumors.

Species:

Homo sapiens, human

Tissue:

Colon

Disease:

Colorectal adenocarcinoma

Gender:

Female, 44 years adult, Caucasian

Morphology:

Epitheloid cells growing as monolayers and in large colonies

Growth Mode:

Adherent

Doubling Time:

40~60hrs

DNA Profile:

Amelogenin: X
CSF1PO: 11,12
D13S317: 11,12/11, 11
D16S539: 11,12
D5S818: 11,12
D7S820: 10
THO1: 6,9
TPOX: 8,9
vWA: 17,19
Cobioer’s Cell Line Authentication Service

Culture Medium:

McCoy's 5a+10% FBS
We strongly suggest to purchase the complete medium from Cobioer.

Cryopreservation medium:

90% Complete Medium+10%DMSO

Antigen Expression:

Blood Type A; Rh+; HLA A1, A3, B12, B17, Cw5

Receptor Expression:

Human adrenergic alpha2A, urokinase receptor (u-PAR), vitamin D (moderate expression), urokinase receptor (u-PAR); vitamin D (moderate expression), human adrenergic alpha2A

Oncogene:

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

Genes Expressed:

Secretory component of IgA; carcinoembryonic antigen (CEA); transforming growth factor beta binding protein; mucin,myc +; ras +; myb +; fos +; sis +; p53 +; abl -; ros -; src -,Blood Type A; Rh+; HLA A1, A3, B12, B17, Cw5,HT-29 cells are negative for CD4, but there is cell surface expression of galactose ceramide (a possible alternative receptor for HIV).

Cellular Products:

Secretory component of IgA; carcinoembryonic antigen (CEA); transforming growth factor beta binding protein; mucin

Tumor Formation:

Yes, in nude mice; forms well differentiated adenocarcinoma consistent with colonic primary (grade I); tumors also form in steroid treated hamsters

Immunology:

Cytokeratin +, cytokeratin-7 -, cytokeratin-8 +, cytokeratin-17 -, cytokeratin-18 +, cytokeratin-19 +, desmin -, endothel -, EpCAM +, GFAP -, HMB-45 -, neurofilament -, vimentin -

Photo:

  

Comments:

Ultrastructural features reported for HT-29 cells include microvilli, microfilaments, large vacuolated mitochondria with dark granules, smooth and rough endoplasmic reticulum with free ribosomes, lipid droplets, few primary and many secondary lysosomes.
The cells express urokinase receptors, but do not have detectable plasminogen activator activity [PubMed ID: 8381394]. HT-29 cells are negative for CD4, but there is cell surface expression of galactose ceramide (a possible alternative receptor for HIV).
The line is positive for expression of c-myc, K-ras, H-ras, N-ras, Myb, sis and fos oncogenes. The p53 antigen is overproduced, and there is a G -> A mutation in codon 273 of the p53 gene resulting in an Arg -> His substitution. N-myc oncogene expression was not detected.
There is a G -> A mutation in codon 273 of the p53 gene resulting in an Arg -> His substitution.

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 1:3 to 1:4 is recommended
Medium Renewal: 2 to 3 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

BRAF

Heterozygous

c.1799T>A

p.V600E

PIK3CA

Heterozygous

c.1345C>A

p.P449T

SMAD4

Homozygous

c.931C>T

p.Q311*

TP53

Homozygous

c.818G>A

p.R273H

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

Expression:

GJC2, CHRAC1, TM4SF4, ST3GAL1
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=19372543; DOI=10.1158/1535-7163.MCT-08-0921; Lorenzi P.L., Reinhold W.C., Varma S., Hutchinson A.A., Pommier Y., Chanock S.J., Weinstein J.N.; "DNA fingerprinting of the NCI-60 cell line panel."; Mol. Cancer Ther. 8:713-724(2009).

2.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).

3.PubMed=19941903; DOI=10.1016/j.jviromet.2009.11.022; Karger A., Bettin B., Lenk M., Mettenleiter T.C.; "Rapid characterisation of cell cultures by matrix-assisted laser desorption/ionisation mass spectrometric typing."; J. Virol. Methods 164:116-121(2010).

4.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).
5.PubMed=22336246; DOI=10.1016/j.bmc.2012.01.017; Kong D., Yamori T.; "JFCR39, a panel of 39 human cancer cell lines, and its application in the discovery and development of anticancer drugs."; Bioorg. Med. Chem. 20:1947-1951(2012).
6.PubMed=22940288; DOI=10.1016/j.jsbmb.2012.08.003; Hobaus J., Fetahu I.S.H., Khorchide M., Manhardt T., Kallay E.; "Epigenetic regulation of the 1,25-dihydroxyvitamin D3 24-hydroxylase (CYP24A1) in colon cancer cells."; J. Steroid Biochem. Mol. Biol. 136:296-299(2013).
7.PubMed=23856246; DOI=10.1158/0008-5472.CAN-12-3342; Abaan O.D., Polley E.C., Davis S.R., Zhu Y.J., Bilke S., Walker R.L., Pineda M., Gindin Y., Jiang Y., Reinhold W.C., Holbeck S.L., Simon R.M., Doroshow J.H., Pommier Y., Meltzer P.S.; "The exomes of the NCI-60 panel: a genomic resource for cancer biology and systems pharmacology."; Cancer Res. 73:4372-4382(2013).
8.PubMed=23933261; DOI=10.1016/j.celrep.2013.07.018; Moghaddas Gholami A., Hahne H., Wu Z., Auer F.J., Meng C., Wilhelm M., Kuster B.; "Global proteome analysis of the NCI-60 cell line panel."; Cell Rep. 4:609-620(2013).
9.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).
10.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).
11.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).
12.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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