生長狀態(tài)：貼壁生長
年限：6 weeks
運(yùn)輸方式：凍存運(yùn)輸
細(xì)胞形態(tài)：上皮樣
品系：BALB/c
細(xì)胞類型：其他細(xì)胞類型
ATCC Number：CRL-1995?
是否是腫瘤細(xì)胞：0
物種來源：小鼠
數(shù)量：大量
器官來源：肝
規(guī)格：20ml Designations: H2.35
Depositors: ?KS Zaret
Biosafety Level:2 [CELLS CONTAIN PAPOVAVIRUS ]
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism: Mus musculus
Morphology:epithelial
Source: Organ: liver
Strain: BALB/c
Cell Type: hepatocyte; SV40 transformed
Permits/Forms:In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Virus Susceptibility:Semliki Forest virus
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.
Tumorigenic:Yes
Age: 6 weeks
Gender: female
Comments: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 33C, and infected with tsA255 (a temperature-sensitive mutant of SV40 virus).
H2.35 cells were cloned from a colony that arose from the SV40 transformed cells.
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.
This cell line provides a model system for studying hepatocyte differentiation in which various hepatocyte phenotypes are inducible.
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.
Propagation: ATCC 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%
Atmosphere: air, 90%; carbon dioxide (CO2), 10%
Temperature: 33.0°C
Subculturing: Protocol:
- Remove and discard culture medium.
- 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.
- 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. - 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 33?C.
Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:10 is recommended
Medium Renewal: 2 to 3 times per week
Preservation: Freeze medium: Complete growth medium supplemented with an additional 20% fetal bovine serum and 5% DMSO
Storage temperature: liquid nitrogen vapor phase
Related Products:recommended serum:ATCC 30-2020
References: 22577: 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
23241: 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
23242: DiPersio CM, et al. The extracellular matrix coordinately modulates liver transcription factors and hepatocyte morphology. Mol. Cell. Biol. 11: 4405-4414, 1991. PubMed: 1875930
23308: Zaret KS, et al. Conditional enhancement of liver-specific gene transcription. Proc. Natl. Acad. Sci. USA 85: 9076-9080, 1988. PubMed: 3194409
