HEP G2/2.2.1 (ATCC® CRL-11997)

Organism: Homo sapiens, human  /  Tissue: liver  /  Disease: hepatocellular carcinoma

Organism Homo sapiens, human
Tissue liver
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease hepatocellular carcinoma
Age 15 years
Gender male
Ethnicity Caucasian
Storage Conditions liquid nitrogen vapor phase
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.
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.
  1. Remove and discard culture medium.
  2. 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.
  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 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.
  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: Every 2 to 3 days
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
Growth Conditions: These cells are slow to attach after subculture. Allow 4 to 5 days for reattachment.
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
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

Basic Documentation
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