• 名称: Hep3B(人肝癌细胞)
  • 货号: CBP60197
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CBP600197

 

 

I. General information

Synonyms:

Hep 3B2_1-7; Hep 3B2; HEP-3B2; HEP3B2; Hep-3B; Hep3B; HEP3B; HEP-3B; Hep 3B

Background:

Established from the tumor tissue of an 8-year-old black boy in 1976; description: cells contain a 2.3 kb integrated hepatitis B virus genome fragment; however there is currently no evidence that this cell line produces infectious Hepatitis B virus. The cells should be handled under laboratory containment level 1. Cells were described to produce a variety of proteins, e.g. alpha-fetoprotein, albumin, transferrin, alpha2-macroglobulin, alpha1-antitrypsin, haptoglobin and others; patented cell line.

Species:

Homo sapiens, human

Tissue:

Liver

Disease:

Hepatocellular carcinoma

Gender:

Male, 8 years juvenile,Black

Morphology:

Epithelial-like cells growing as monolayers

Growth Mode:

Adherent

Doubling Time:

40~50hrs

DNA Profile:

Amelogenin: X
CSF1PO: 8
D13S317: 12,14
D16S539: 10
D5S818: 13
D7S820: 8,10
THO1: 6,7
TPOX: 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:

 

Genes Expressed:

Alpha fetoprotein (alpha-fetoprotein); hepatitis B surface antigen (HBsAg); albumin; alpha2 macroglobulin (alpha-2-macroglobulin); alpha1 antitrypsin (alpha-1-antitrypsin); transferrin;,alpha1 antichymotrypsin (alpha-1-antichymotrypsin); haptoglobin; ceruloplasmin; plasminogen; complement (C3, 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:

Alpha fetoprotein (alpha-fetoprotein); hepatitis B surface antigen (HBsAg); albumin; alpha2 macroglobulin (alpha-2-macroglobulin); alpha1 antitrypsin (alpha-1-antitrypsin); transferrin;
alpha1 antichymotrypsin (alpha-1-antichymotrypsin); haptoglobin; ceruloplasmin; plasminogen; complement (C3, 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); Gc globulin

Effects:

Yes, forms tumors in nude mice

Immunology:

cytokeratin +, cytokeratin-7 (+), cytokeratin-8 +, cytokeratin-17 -, cytokeratin-18 +, desmin -, endothel -, GFAP -, neurofilament -, vimentin +

Comments:

The cell line contains parts of the hepatitis B virus (HBV) genome integrated into the eukaryotic genome. The HBV surface antigen (HBsAg) is expressed, but no active viruses are produced by the cells. The cells can thus be handled under biosafety level 1.

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

NA

NA

NA

NA

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=12029633; DOI=10.1053/jhep.2002.33683; Yasui K., Arii S., Zhao C., Imoto I., Ueda M., Nagai H., Emi M., Inazawa J.; "TFDP1, CUL4A, and CDC16 identified as targets for amplification at 13q34 in hepatocellular carcinomas."; Hepatology 35:1476-1484(2002).

2.PubMed=15767549; DOI=10.1158/1535-7163.MCT-04-0234; Nakatsu N., Yoshida Y., Yamazaki K., Nakamura T., Dan S., Fukui Y., Yamori T.; "Chemosensitivity profile of cancer cell lines and identification of genes determining chemosensitivity by an integrated bioinformatical approach using cDNA arrays."; Mol. Cancer Ther. 4:399-412(2005).

3.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).

4.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).
5.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).

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