H2.35 (ATCC® CRL-1995)

Organism: Mus musculus, mouse  /  Cell Type: hepatocyte; SV40 transforme  /  Tissue: liver  / 

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Organism Mus musculus, mouse
Tissue
liver
Cell Type hepatocyte; SV40 transforme
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 [CELLS CONTAIN PAPOVAVIRUS]
Age 6 weeks
Gender female
Strain BALB/c
Applications
The line is a good transfection recipient for assaying the function of genetic regulatory sequences for various liver genes that do not function in other hepatocyte-derived cell lines.
This cell line provides a model system for studying hepatocyte differentiation in which various hepatocyte phenotypes are inducible.
Storage Conditions liquid nitrogen vapor phase
Derivation
H2.35 cells were cloned from a colony that arose from the SV40 transformed cells.
H2.35 is an epithelial-like cell line derived from a primary hepatocyte culture.
Cells were collected from the livers of 6 week old female BALB/c mice, incubated at 33°C, and infected with tsA255 (a temperature-sensitive mutant of SV40 virus).
Clinical Data
Cells were collected from the livers of 6 week old female BALB/c mice, incubated at 33°C, and infected with tsA255 (a temperature-sensitive mutant of SV40 virus).
female
Tumorigenic Yes
Effects
Yes, in BALB/c nu/nu mice
Virus Susceptibility Semliki Forest virus
Comments
The cells undergo morphological changes in response to extracellular matrix gel substrata, and albumin gene transcription is induced.
Culturing the cells in serum-free medium and on collagen gels causes the induction of several liver enriched transcription factors.
Complete Growth Medium Dulbecco's modified Eagle's medium with 1.0 g/L glucose and 200 nM dexamethasone, 96.0%; fetal bovine serum, 4.0%
Subculturing
Protocol:
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.03% 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 33°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 33°C.
Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:10 is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation
Freeze medium: Complete growth medium supplemented with an additional 20% fetal bovine serum and 5% DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 90%; carbon dioxide (CO2), 10%
Temperature: 33°C
Temperature Effects
Restrictive temperature: 39° C expresses 100-fold higher levels of albumin mRNA
Permissive temperature: 33°C express only very low levels of albumin mRNA
The cells are temperature sensitive for the expression of serum albumin mRNA.
Name of Depositor KS Zaret
References

Liu JK, et al. Extracellular signals that regulate liver transcription factors during hepatic differentiation in vitro. Mol. Cell. Biol. 11: 773-784, 1991. PubMed: 1990282

Zaret KS, Stevens KA. Selection and analysis of galactose metabolic pathway variants of a mouse liver cell line. Mol. Cell. Biol. 10: 4582-4589, 1990. PubMed: 2167434

DiPersio CM, et al. The extracellular matrix coordinately modulates liver transcription factors and hepatocyte morphology. Mol. Cell. Biol. 11: 4405-4414, 1991. PubMed: 1875930

Zaret KS, et al. Conditional enhancement of liver-specific gene transcription. Proc. Natl. Acad. Sci. USA 85: 9076-9080, 1988. PubMed: 3194409

The cells are temperature sensitive for the expression of serum albumin mRNA.

Expression of albumin mRNA requires growth on serum-free medium and collagen-coated surface.

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

Liu JK, et al. Extracellular signals that regulate liver transcription factors during hepatic differentiation in vitro. Mol. Cell. Biol. 11: 773-784, 1991. PubMed: 1990282

Zaret KS, Stevens KA. Selection and analysis of galactose metabolic pathway variants of a mouse liver cell line. Mol. Cell. Biol. 10: 4582-4589, 1990. PubMed: 2167434

DiPersio CM, et al. The extracellular matrix coordinately modulates liver transcription factors and hepatocyte morphology. Mol. Cell. Biol. 11: 4405-4414, 1991. PubMed: 1875930

Zaret KS, et al. Conditional enhancement of liver-specific gene transcription. Proc. Natl. Acad. Sci. USA 85: 9076-9080, 1988. PubMed: 3194409

The cells are temperature sensitive for the expression of serum albumin mRNA.

Expression of albumin mRNA requires growth on serum-free medium and collagen-coated surface.