• 名称: HuH-7(人肝癌细胞)
  • 货号: CBP60202
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CBP600202

 

 

I. General information

Synonyms:

HuH-7; HUH-7; HuH7; Huh7; HUH7; JTC-39

Background:

NA

Species:

Homo sapiens, human

Tissue:

Liver, gallbladder

Disease:

Well differentiated hepato cellular carcinoma.

Gender:

Male, 57-year-old

Morphology:

Epithelial-like

Growth Mode:

Adherent

Doubling Time:

Approx. 37 hrs

DNA Profile:

Amelogenin: X,X
CSF1PO: 11,11
D13S317: 10,10/10,11
D16S539: 10,10
D5S818: 12,12
D7S820: 11,11
THO1: 7,7
TPOX: 8,11
vWA: 16,18/18,18
Cobioer’s Cell Line Authentication Service

Culture Medium:

DMEM+10%FB

We strongly suggest to purchase the complete medium from Cobioer.

Cryopreservation medium:

90% Complete Medium+10%DMSO

Photo:

 

Viruses:

CMV(-), EBV(-), HHV6(-), HHV7(-), BKJ(-), JCV(-), ADV1(-), parvoB19(-), HBV(-), HTLV1(-), HTLV2(-), HIV1(-), HIV2(-), HPV18(nt) [-/negative, +/positive, nt/not tested]

Comments:

This cell line can be grown in serum-free medium. Many serum proteins with alpha phyto protein are released into the medium.

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 1:2 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

ADAMTSL3

Unknown

c.C>T

p.R1005C

CDK13

Unknown

c.G>T

p.Q544H

MAP3K1

Unknown

c.CAA>-

p.T949del

NOTCH1

Unknown

c.T>C

p.Q184R

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

Expression:

AQP10, RHBG, AMT
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=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).

2. PubMed=22460905; DOI=10.1038/nature11003; Barretina J., Caponigro G., Stransky N., Venkatesan K., Margolin A.A., Kim S., Wilson C.J., Lehar J., Kryukov G.V., Sonkin D., Reddy A., Liu M., Murray L., Berger M.F., Monahan J.E., Morais P., Meltzer J., Korejwa A., Jane-Valbuena J., Mapa F.A., Thibault J., Bric-Furlong E., Raman P., Shipway A., Engels I.H., Cheng J., Yu G.K., Yu J., Aspesi P. Jr., de Silva M., Jagtap K., Jones M.D., Wang L., Hatton C., Palescandolo E., Gupta S., Mahan S., Sougnez C., Onofrio R.C., Liefeld T., MacConaill L., Winckler W., Reich M., Li N., Mesirov J.P., Gabriel S.B., Getz G., Ardlie K., Chan V., Myer V.E., Weber B.L., Porter J., Warmuth M., Finan P., Harris J.L., Meyerson M., Golub T.R., Morrissey M.P., Sellers W.R., Schlegel R., Garraway L.A.; "The Cancer Cell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity."; Nature 483:603-607(2012).

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=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=25574106; DOI=10.3748/wjg.v21.i1.311; Cevik D., Yildiz G., Ozturk M.; "Common telomerase reverse transcriptase promoter mutations in hepatocellular carcinomas from different geographical locations."; World J. Gastroenterol. 21:311-317(2015).
7. PubMed=27397505; DOI=10.1016/j.cell.2016.06.017; Iorio F., Knijnenburg T.A., Vis D.J., Bignell G.R., Menden M.P., Schubert M., Aben N., Goncalves E., Barthorpe S., Lightfoot H., Cokelaer T., Greninger P., van Dyk E., Chang H., de Silva H., Heyn H., Deng X., Egan R.K., Liu Q., Mironenko T., Mitropoulos X., Richardson L., Wang J., Zhang T., Moran S., Sayols S., Soleimani M., Tamborero D., Lopez-Bigas N., Ross-Macdonald P., Esteller M., Gray N.S., Haber D.A., Stratton M.R., Benes C.H., Wessels L.F.A., Saez-Rodriguez J., McDermott U., Garnett M.J.; "A landscape of pharmacogenomic interactions in cancer."; Cell 166:740-754(2016).

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