| 產(chǎn)品名稱 |
HEP G2/2.2.1 |
| 商品貨號 |
B164594 |
| Organism |
Homo sapiens, human |
| Tissue |
liver |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Disease |
hepatocellular carcinoma |
| Age |
15 years |
| Gender |
male |
| Ethnicity |
Caucasian |
| Storage Conditions |
liquid nitrogen vapor phase |
| Disclosure |
This material is cited in a US or other Patent and may not be used to
infringe the claims. Depending on the wishes of the Depositor, ATCC may be
required to inform the Patent Depositor of the party to which the material was
furnished. This material may not have been produced or characterized by ATCC. |
| Derivation |
Cell line was derived from the hepatocellular carcinoma cell line, HepG2 (ATCC HB-8065). The parental cells were stably transfected at passage 48 with a human cholesterol 7 alpha-hydroxylase (CYP7) minigene/Luciferase construct.
|
| Clinical Data |
15 years Caucasian male |
| Comments |
The vector map shown in U.S. Patent 5,821,057 notes the luciferase reporter is under the control of a human cholesterol 7-alpha-hydroxylase (CYP7) regulatory minigene. The cells must be treated with a test compound that up-regulates expression of the minigene and luciferase. Additional information regarding the transfection and selection of these cells is provided in the depositor's publications, including US Patent 5,821,057.
(Chiang JY, Stroup D. Assay for agents that affect cholesterol 7alpha-hydroxylase expression and a characterization of its regulatory elements. US Patent 5,821,057 dated Oct 13 1998) |
| Complete Growth Medium |
A 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium with 2.5 mM L-glutamine, 15 mM HEPES, 0.5 mM sodium pyruvate and 1200 mg/L sodium bicarbonate and supplemented with 0.4 mg/ml G418, 90%; fetal bovine serum, 10%
|
| Subculturing |
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
- Remove and discard culture medium.
-
Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
-
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 5 to 15 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.
-
Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
-
Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended
Medium Renewal: Every 2 to 3 days
Note: These cells are slow to attach after subculture. Allow 4 to 5 days for reattachment. |
| Cryopreservation |
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions |
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C |
| STR 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 |
| Name of Depositor |
Northeastern Ohio University College of Medicine |
| Deposited As |
human |
| U.S. Patent Number |
|
| Passage History |
Cell line was derived from the hepatocellular carcinoma cell line, HepG2 (ATCC HB-8065). The parental cells were stably transfected at passage 48 with a human cholesterol 7 alpha-hydroxylase (CYP7) minigene/Luciferase construct. |
| References |
Chiang JY, Stroup D. Assay for agents that affect cholesterol 7alpha-hydroxylase expression and a characterization of its regulatory elements. US Patent 5,821,057 dated Oct 13 1998
|
