• 名称: HepG2(人肝癌细胞)
  • 货号: CBP60199
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CBP600199

 

 

I. General information

Synonyms:

HEP-G2; Hep G2; HEP G2; HepG2; HEPG2

Background:

Established from the tumor tissue of a 15-year-old Argentine boy with well differentiated hepatocellular carcinoma in 1975; cell line is patented; cells were described to not harbor a hepatitis B virus genome; cells reportedly produce a variety of proteins: alpha-fetoprotein, albumin, alpha2-macroglobulin, alpha1-antitrypsin, transferrin, alpha1-antichymotrypsin, haptoglobin, ceruloplasmin, plasminogen, complement (C3, C4), C3 activator, fibrinogen, alpha1-acid glycoprotein, alpha2-HS glycoprotein, ß-lipoprotein, retinol binding protein. They have been grown successfully in large scale cultivation systems. The cells will respond to stimulation with human growth hormone.

Species:

Homo sapiens, human

Tissue:

Liver

Disease:

Hepatocellular carcinoma

Gender:

Male, 15 years, Caucasian

Morphology:

Epithelial-like cells growing as monolayers and in small aggregates

Growth Mode:

Adherent

Doubling Time:

50~60hrs

DNA Profile:

Amelogenin: X,Y
CSF1PO: 10,11
D13S317: 9,13
D16S539: 12,13
D5S818: 11,12
D7S820: 10
THO1: 9
TPOX: 8,9
vWA: 17
Cobioer’s Cell Line Authentication Service

Culture Medium:

MEM+10%FBS+1%NEAA+1%NaP
We strongly suggest to purchase the complete medium from Cobioer.

Cryopreservation medium:

90% Complete Medium+10%DMSO

Photo:

 

Receptor Expression:

Insulin; insulin-like growth factor II (IGF II)

Genes Expressed:

Alpha-fetoprotein (alpha fetoprotein); albumin; alpha2 macroglobulin (alpha-2-macroglobulin); alpha1 antitrypsin (alpha-1-antitrypsin); transferrin; alpha1 antichymotrypsin; (alpha-1-antichymotrypsin); haptoglobin; ceruloplasmin; plasminogen;,complement (C4); C3 activator; fibrinogen; alpha1 acid glycoprotein (alpha-1 acid glycoprotein); alpha2 HS glycoprotein (alpha-2-HS-glycoprotein); beta lipoprotein (beta-lipoprotein); retinol binding protein (retinol-binding protein)

Cellular Products:

Cellular products: alpha-fetoprotein (alpha fetoprotein); albumin; alpha2 macroglobulin (alpha-2-macroglobulin); alpha1 antitrypsin (alpha-1-antitrypsin); transferrin; alpha1 antichymotrypsin; (alpha-1-antichymotrypsin); haptoglobin; ceruloplasmin; plasminogen;

complement (C4); C3 activator; fibrinogen; alpha1 acid glycoprotein (alpha-1 acid glycoprotein); alpha2 HS glycoprotein (alpha-2-HS-glycoprotein); beta lipoprotein (beta-lipoprotein); retinol binding protein (retinol-binding protein)

Effects:

No, in immunosuppressed mice

Yes, in semisolid medium

Immunology:

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

Comments:

The cells express 3-hydroxy-3-methylglutaryl-CoA reductase and hepatic triglyceride lipase activities.
The cells demonstrate decreased expression of apoA-I mRNA and increased expression of catalase mRNA in response to gramoxone (oxidative stress).

There is no evidence of a Hepatitis B virus genome in this cell line.

Misidentified: Originally thought to be a hepatocellular carcinoma cell line but shown to be from an hepatoblastoma.

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: 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

NRAS

Unknown

c.182A>T

p.Q61L

RET

Unknown

c.G>A

p.D567N

ATM

Unknown

c.G>A

p.V2906I

PRKDC

Unknown

c.->G

p.L1244fs

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=19751877; DOI=10.1016/j.humpath.2009.07.003; Lopez-Terrada D., Cheung S.W., Finegold M.J., Knowles B.B.; "Hep G2 is a hepatoblastoma-derived cell line."; Hum. Pathol. 40:1512-1515(2009).

2.PubMed=20069059; DOI=10.1155/2010/437143; Srisomsap C., Sawangareetrakul P., Subhasitanont P., Chokchaichamnankit D., Chiablaem K., Bhudhisawasdi V., Wongkham S., Svasti J.; "Proteomic studies of cholangiocarcinoma and hepatocellular carcinoma cell secretomes."; J. Biomed. Biotechnol. 2010:437143-437143(2010).

3.PubMed=20228232; DOI=10.1124/dmd.109.031831;Hart S.N., Li Y., Nakamoto K., Subileau E.-A., Steen D., Zhong X.-B.;"A comparison of whole genome gene expression profiles of HepaRG cells and HepG2 cells to primary human hepatocytes and human liver tissues.";Drug Metab. Dispos. 38:988-994(2010).

4.PubMed=22278370; DOI=10.1074/mcp.M111.014050;Geiger T., Wehner A., Schaab C., Cox J., Mann M.;"Comparative proteomic analysis of eleven common cell lines reveals ubiquitous but varying expression of most proteins.";Mol. Cell. Proteomics 11:M111.014050-M111.014050(2012).
5.PubMed=23325432; DOI=10.1101/gr.147942.112;Varley K.E., Gertz J., Bowling K.M., Parker S.L., Reddy T.E., Pauli-Behn F., Cross M.K., Williams B.A., Stamatoyannopoulos J.A., Crawford G.E., Absher D.M., Wold B.J., Myers R.M.; "Dynamic DNA methylation across diverse human cell lines and tissues."; Genome Res. 23:555-567(2013).
6.PubMed=23505090; DOI=10.1002/hep.26402;Wang K., Lim H.Y., Shi S., Lee J., Deng S., Xie T., Zhu Z., Wang Y., Pocalyko D., Yang W.J., Rejto P.A., Mao M., Park C.-K., Xu J.; "Genomic landscape of copy number aberrations enables the identification of oncogenic drivers in hepatocellular carcinoma."; Hepatology 58:706-717(2013).
7.PubMed=23887712; DOI=10.1038/ncomms3218; Nault J.-C., Mallet M., Pilati C., Calderaro J., Bioulac-Sage P., Laurent C., Laurent A., Cherqui D., Balabaud C., Zucman-Rossi J.; "High frequency of telomerase reverse-transcriptase promoter somatic mutations in hepatocellular carcinoma and preneoplastic lesions."; Nat. Commun. 4:2218-2218(2013).
8.PubMed=24116068; DOI=10.1371/journal.pone.0075692; Weiskirchen R., Weimer J., Meurer S.K., Kron A., Seipel B., Vater I., Arnold N., Siebert R., Xu L., Friedman S.L., Bergmann C.; "Genetic characteristics of the human hepatic stellate cell line LX-2."; PLoS ONE 8:E75692-E75692(2013).
9.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).
10.PubMed=26825538; DOI=10.1016/j.jprot.2016.01.016;Wisniewski J.R., Vildhede A., Noren A., Artursson P.; "In-depth quantitative analysis and comparison of the human hepatocyte and hepatoma cell line HepG2 proteomes."; J. Proteomics 136:234-247(2016).

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